Use of clostridial neurotoxin variants for treating neurological disorders

Abstract

The present invention relates to a polypeptide for use in promoting neuronal growth or neuronal repair to treat a neurological disorder in a subject, wherein the polypeptide comprises: a clostridial neurotoxin light chain (L-chain) or fragment thereof; and / or a fragment of a clostridial neurotoxin heavy chain (H-chain). Other polypeptides for promoting neuronal growth or neuronal repair to treat neurological disorders in a subject, as well as corresponding methods and uses, are also provided.

Description

[0001] This application is a divisional application of Chinese application 202080068881.4, filed on September 30, 2020, entitled "Use of Clostridium neurotoxin variants for the treatment of neurological disorders".

[0002] This invention relates to the treatment of neurological disorders.

[0003] Neurological disorders include neuronal damage, neurodegenerative disorders, sensory disorders, and autonomic nervous system disorders.

[0004] Neuronal injury, such as spinal cord injury (SCI), leads to the degeneration of damaged axons, thereby preventing normal sensory, motor, and autonomic nervous functions. Recovery can occur through endogenous mechanisms, such as the regeneration of damaged axons and the lateral column sprouting of undamaged axons, resulting in the restoration of innervation to the denervated target. However, the regenerative capacity of damaged neurons (especially in the spinal cord) is limited in adult mammals, and patients may suffer from various disabilities, which significantly impacts their quality of life.

[0005] Conventional treatments for neuronal injury include interleukin-6 (IL-6) and stem cell transplantation; however, few are in the development stage for clinical use. Therefore, there remains a need for therapeutic agents for neuronal injury that can promote neuronal growth or repair.

[0006] Bacteria in the genus *Clostridia* produce highly toxic and specific protein toxins that poison the neurons and other cells to which they are delivered. Examples of such clostridial toxins include the neurotoxin TeNT produced by *C. tetanus* and the neurotoxin BoNT serotypes AG and X produced by *C. botulinum* (see WO2018 / 009903 A2), as well as the neurotoxins produced by *C. baratii* and *C. butyricum*.

[0007] Among clostridial neurotoxins, some are known to be the most potent. For example, depending on serotype, botulinum neurotoxin has a median lethal dose (LD50) in mice ranging from 0.5 to 5 ng / kg. 50 The values ​​are as follows. Both tetanus toxin and botulinum toxin work by inhibiting the function of affected neurons, particularly by inhibiting the release of neurotransmitters. Botulinum toxin acts on the neuromuscular junction and inhibits cholinergic transmission in the peripheral nervous system, while tetanus toxin acts on the central nervous system.

[0008] In nature, clostridium neurotoxins are synthesized as single-chain polypeptides, which are modified post-translational by proteolytic cleavage events to form two polypeptide chains linked together by disulfide bonds. Cleavage occurs at specific cleavage sites, often called activation sites, located between cysteine ​​residues that provide the interchain disulfide bonds. This double-chain form is the active form of the toxin. These two chains are called the heavy chain (H-chain) (with a molecular weight of approximately 100 kDa) and the light chain (L-chain) (with a molecular weight of approximately 50 kDa). The H-chain contains the N-terminal translocation component (H... N (structural domain) and C-terminal targeting component (H) C (Structural domain). The cleavage site is located between the L-chain and the translocation structural domain components. In H C After the domain binds to its target neuron and internalizes the bound toxin into the cell via endosomes, H... N The domain translocates the L-chain across the endosome membrane and into the cytosol, while the L-chain provides protease function (also known as a non-cytotoxic protease).

[0009] Non-cytotoxic proteases exert their effects by proteolytically cleaving intracellular transport proteins called SNARE proteins (such as SNAP-25, VAMP, or synaptic fusion proteins). The acronym SNARE originates from the term soluble NSF attachment receptor (NSF receptor). S oluble N SF A ttachment Re ceptor), where NSF represents N-ethylmaleimide sensitizer ( N -ethylmaleimide- S ensitive F SNARE proteins are components of intracellular vesicle fusion bodies and are therefore secreted molecules via the transport of vesicles from the cell. The protease function is a zinc-dependent endopeptidase activity, exhibiting high substrate specificity for SNARE proteins. Therefore, once delivered to the desired target cell, the non-cytotoxic protease is able to inhibit cellular secretion from the target cell. The L-chain protease of clostridium neurotoxin is a non-cytotoxic protease that cleaves SNARE proteins.

[0010] Given the ubiquity of SNARE proteins, clostridial neurotoxins (such as botulinum toxin) have been successfully used in a variety of treatments.

[0011] WO 2016 / 170501 A1 describes the use of catalytically active full-length BoNT / A (containing L-chains and complete H-chains, including H+). N and H CBoNT / A is used to treat paralysis caused by spinal cord injury. WO 2016 / 170501A1 teaches that each functional domain of BoNT / A is essential for the observed therapeutic effect, including H-chain binding and translocation capabilities and L-chain non-cytotoxic protease activity. As mentioned above, full-length clostridium neurotoxins are highly effective, but specific safety procedures are required when handling the toxin. Furthermore, the spread of the toxin from the target tissue is considered a cause of potentially life-threatening adverse side effects in extreme cases. This may be a particularly concerning issue when using clostridium neurotoxin therapeutics (such as BoNT therapeutics) at high doses, concentrations, and injections. Adverse effects reported with this problem associated with commercial BoNT / A therapies include weakness, generalized muscle weakness, diplopia, ptosis, dysphagia, dysarthria, dysarthria, urinary incontinence, and respiratory distress. Dysphagia and respiratory distress can be life-threatening, and deaths have been reported related to the spread of toxin effects. Therefore, a safer therapeutic agent is needed to promote neuronal growth or repair.

[0012] Given their size, the use of the full-length Clostridium neurotoxin (~150 kDa) or its intact H-chain (~100 kDa) is associated with an increased risk of inducing an immune response in subjects treated with the peptide. Furthermore, the entire H-chain (especially H...) C The presence of the structural domain allows the peptide to bind to the target receptor of clostridium neurotoxin, which may be associated with undesirable off-target effects in subjects who have been given the peptide.

[0013] The present invention overcomes one or more of the above-mentioned problems.

[0014] The inventors of this invention have surprisingly discovered fragments containing clostridium neurotoxin L-chain and / or clostridium neurotoxin H-chain (e.g., translocation domains (H... N ) or receptor-binding domain (H C The peptides of [the drug] promote neuronal growth or repair, thus revealing their utility in treating neurological disorders. Advantageously, this allows for the use of non-toxic (or substantially non-toxic) fragments of clostridium neurotoxins, which are smaller in size (compared to full-length H-chains or full-length clostridium neurotoxins) and less likely to elicit an immune response in subjects administered the fragments. Furthermore, the non-toxic (or substantially non-toxic) fragments are less expensive to manufacture and / or simpler to produce compared to full-length clostridium neurotoxins. Additionally, the non-toxic (or substantially non-toxic) fragments constitute more defined therapeutic agents than full-length clostridium neurotoxins, and given the shorter peptide length, the possibility of cysteine ​​shuffling between domains is reduced.

[0015] Therefore, in one aspect, the present invention provides a polypeptide for promoting neuronal growth or neuronal repair to treat neurological disorders in a subject, wherein the polypeptide comprises:

[0016] Clostridium neurotoxin light chain (L-chain) or fragments thereof; and / or

[0017] Fragments of the clostridium neurotoxin heavy chain (H-chain).

[0018] In a related aspect, a method for promoting neuronal growth or repair to treat neurological disorders in a subject is provided, the method comprising administering a peptide to the subject, wherein the peptide comprises:

[0019] Clostridium neurotoxin L-chain or fragments thereof; and / or

[0020] Fragments of the H-chain of Clostridium neurotoxin.

[0021] In another aspect, the use of the polypeptide in the preparation of a medicament for promoting neuronal growth or neuronal repair to treat neurological disorders in a subject is provided, wherein the polypeptide comprises:

[0022] Clostridium neurotoxin L-chain or fragments thereof; and / or

[0023] Fragments of the H-chain of Clostridium neurotoxin.

[0024] In one aspect, the present invention provides a polypeptide for treating a neurological disorder in a subject, wherein the polypeptide comprises:

[0025] Clostridium neurotoxin light chain (L-chain) or fragments thereof; and / or

[0026] Fragments of the clostridium neurotoxin heavy chain (H-chain).

[0027] In related aspects, a method for treating a neurological disorder in a subject is provided, the method comprising administering a polypeptide to the subject, wherein the polypeptide comprises:

[0028] Clostridium neurotoxin L-chain or fragments thereof; and / or

[0029] Fragments of the H-chain of Clostridium neurotoxin.

[0030] In another aspect, the use of the polypeptide in the preparation of a medicament for treating a neurological disorder in a subject is provided, wherein the polypeptide comprises:

[0031] Clostridium neurotoxin L-chain or fragments thereof; and / or

[0032] Fragments of the H-chain of Clostridium neurotoxin.

[0033] In one embodiment, the polypeptide of the present invention comprises a clostridium neurotoxin L-chain. Preferably, the L-chain is non-catalytically active.

[0034] Therefore, in one aspect, the present invention provides a polypeptide for promoting neuronal growth or neuronal repair to treat neurological disorders in a subject, wherein the polypeptide comprises a non-catalytically active clostridium neurotoxin L-chain.

[0035] In a related aspect, the present invention provides a method for promoting neuronal growth or neuronal repair to treat neurological disorders in a subject, the method comprising administering a polypeptide to the subject, wherein the polypeptide comprises a non-catalytically active clostridium neurotoxin L-chain.

[0036] In another aspect, the present invention provides the use of a polypeptide comprising a non-catalytically active clostridium neurotoxin L-chain in the preparation of a medicament for promoting neuronal growth or neuronal repair to treat neurological disorders in a subject.

[0037] In one aspect, the present invention provides a polypeptide for treating a neurological disorder in a subject, wherein the polypeptide comprises a non-catalytically active clostridium neurotoxin L-chain.

[0038] In a related aspect, the present invention provides a method for treating a neurological disorder in a subject, the method comprising administering a polypeptide to the subject, wherein the polypeptide comprises a non-catalytically active clostridium neurotoxin L-chain.

[0039] In another related aspect, the present invention provides the use of a polypeptide comprising a non-catalytically active clostridium neurotoxin L-chain in the preparation of a medicament for treating a neurological disorder in a subject.

[0040] The inventors of this invention are the first to demonstrate that the catalytic activity of the L-chain of clostridium neurotoxin is not essential for promoting neuronal growth or neuronal repair. Therefore, this invention allows for the provision of safer (less toxic) therapeutic agents.

[0041] The active clostridium neurotoxin L-chain possesses non-cytotoxic protease activity. Specifically, the active clostridium neurotoxin L-chain exhibits endopeptidase activity and is capable of cleaving proteins of the extracellular fusion apparatus in target cells. The proteins of the extracellular fusion apparatus are preferably SNARE proteins, such as SNAP-25, small synaptic vesicle protein / VAMP, or synaptic fusion proteins.

[0042] As used herein, the term "non-catalytically active" with respect to the L-chain of clostridial neurotoxin means that the L-chain substantially does not exhibit non-cytotoxic protease activity, preferably meaning that the L-chain does not exhibit non-cytotoxic protease activity. In one embodiment, the non-catalytically active clostridial neurotoxin L-chain is a chain that does not cleave proteins of extracellular fusion devices in target cells. The term "substantially non-cytotoxic protease activity" means that the clostridial neurotoxin L-chain has less than 5% of the non-cytotoxic protease activity of the catalytically active clostridial neurotoxin L-chain, for example, less than 2%, 1%, or preferably less than 0.1%. Non-cytotoxic protease activity can be assessed by incubating the test clostridial neurotoxin L-chain with SNARE protein and comparing the amount of SNARE protein cleaved by the test clostridial neurotoxin L-chain with the amount of SNARE protein cleaved by the catalytically active clostridial neurotoxin L-chain under the same conditions. Conventional techniques, such as SDS-PAGE and Western blotting, can be used to quantify the amount of cleaved SNARE protein. Suitable in vitro assays are described in WO 2019 / 145577 A1, which is incorporated herein by reference.

[0043] Cell-based and in vivo assays can also be used to determine whether clostridial neurotoxins containing L-chains and functional cell-binding and translocation domains possess non-cytotoxic protease activity. Assays such as the Digital Abduction Score (DAS), Dorsal Root Ganglion (DRG) assay, Spinal Cord Neuron (SCN) assay, and Mouse Phrenic Nerve Lateral Diaphragm (PNHD) assay are routine in the art. Suitable assays for determining non-cytotoxic protease activity may be those described in Donald et al. (2018), Pharmacol ResPerspect, e00446, 1-14, which are incorporated herein by reference.

[0044] The catalytically inactive L-chain may have one or more mutations that inactivate the catalytic activity. For example, the catalytically inactive BoNT / A L-chain may contain mutations in the active site residues, such as His223, Glu224, His227, Glu262, and / or Tyr366. The position number corresponds to the amino acid position of SEQ ID NO:62 and can be determined by comparing the polypeptide with SEQ ID NO:62. Since the presence of a methionine residue at position 1 of SEQ ID NO:62 is optional, those skilled in the art will consider the presence / absence of the methionine residue when determining the amino acid residue number. For example, if SEQ ID NO:62 includes methionine, the position number will be as defined above (e.g., His223 would be His223 of SEQ ID NO:62). Alternatively, if methionine is absent in SEQ ID NO:62, the amino acid residue number should be changed by -1 (e.g., His223 would be His222 of SEQ ID NO:62). Similar considerations apply when methionine at position 1 is present or absent in other polypeptide sequences described herein, and those skilled in the art will readily determine the correct amino acid residue number using conventional techniques in the art.

[0045] In a particularly preferred embodiment, the polypeptide of the present invention may comprise modified BoNT / A or a fragment thereof (preferably BoNT / AH). C (A structural domain or a fragment thereof). The modified BoNT / A or its fragments may contain modifications at one or more amino acid residues selected from the following: ASN 886, ASN 905, GLN 915, ASN 918, GLU 920, ASN 930, ASN 954, SER 955, GLN 991, GLU 992, GLN 995, ASN 1006, ASN 1025, ASN 1026, ASN 1032, ASN 1043, ASN 1046, ASN 1052, ASP 1058, HIS 1064, ASN 1080, GLU 1081, GLU 1083, ASP 1086, ASN 1188, ASP 1213, GLY 1215, ASN 1216, GLN 1229, ASN 1242, ASN 1243, SER 1274, and THR1277. This modified BoNT / A or fragment thereof can be demonstrated to have fewer or no side effects compared to the use of known BoNT / A. The increased tissue-retention properties of the modified BoNT / A of the present invention can also provide increased potency and / or duration of action, and can allow for a lower dose (or an increased dose without any additional side effects) compared to known clostoxin therapeutics, thus providing further advantages.

[0046] When compared to the unmodified BoNT / A as shown in SEQ ID NO:62, the modification can be a modification, wherein the amino acid residue number is determined by comparison with SEQ ID NO:62. Because the presence of a methionine residue at position 1 of SEQ ID NO:62 (and the SEQ ID NO corresponding to the modified BoNT / A polypeptide or fragment thereof described herein) is optional, those skilled in the art will consider the presence / absence of the methionine residue when determining the amino acid residue number. For example, if SEQ ID NO:62 includes methionine, the position number will be as defined above (e.g., ASN 886 would be ASN 886 of SEQ ID NO:62). Alternatively, if methionine is absent in SEQ ID NO:2, the amino acid residue number should be changed by -1 (e.g., ASN 886 would be ASN 885 of SEQ ID NO:62). Similar considerations apply when methionine is present / absent at position 1 of other polypeptide sequences described herein, and those skilled in the art will readily determine the correct amino acid residue number using techniques conventional in the art.

[0047] The modified amino acid residues are surface-exposed amino acid residues.

[0048] The modified BoNT / A or fragment thereof may contain modifications at one or more amino acid residues selected from the following: ASN 886, ASN 930, ASN 954, SER 955, GLN 991, ASN 1025, ASN 1026, ASN 1052, ASN 1188, ASP 1213, GLY 1215, ASN 1216, GLN 1229, ASN 1242, ASN 1243, SER 1274, and THR 1277.

[0049] When used in the content of a modified BoNT / A or a fragment thereof, the term "one or more amino acid residues" preferably means at least 2, 3, 4, 5, 6, or 7 specified amino acid residues. Therefore, a modified BoNT / A may contain at least 2, 3, 4, 5, 6, or 7 (preferably 7) modifications at specified amino acid residues. A modified BoNT / A or a fragment thereof may contain 1-30, 3-20, or 5-10 amino acid modifications. More preferably, when used in the content of a modified BoNT / A or a fragment thereof, the term "one or more amino acid residues" refers to all specified amino acid residues.

[0050] Preferably, apart from one or more amino acid modifications at the specified amino acid residues, the modified BoNT / A or its fragments do not contain any further amino acid modifications compared to SEQ ID NO:62.

[0051] Modifications selected from:

[0052] i. Replace the amino acid residues exposed on the acidic surface with basic amino acid residues;

[0053] ii. Replace the amino acid residues exposed on the acidic surface with uncharged amino acid residues;

[0054] iii. Replace uncharged surface-exposed amino acid residues with basic amino acid residues;

[0055] iv. Inserting basic amino acid residues; and

[0056] v. To remove amino acid residues exposed on an acidic surface.

[0057] The modifications described above result in modified BoNT / A or fragments thereof having increased positive surface charge and increased isoelectric point compared to the corresponding unmodified BoNT / A or fragments thereof.

[0058] The isoelectric point (pI) is a specific property of a given protein. As is well known in the art, proteins are made up of a specific sequence of amino acids (also called amino acid residues in proteins). Each amino acid in the twenty standard groups has a different side chain (or R group), meaning that each amino acid residue in a protein exhibits different chemical properties, such as charge and hydrophobicity. These properties can be influenced by the surrounding chemical environment, such as temperature and pH. The overall chemical characterization of a protein will depend on the sum of these different factors.

[0059] Some amino acid residues (described in detail below) have ionizable side chains that can exhibit a charge depending on the surrounding pH. At a given pH, whether such a side chain is charged depends on the pKa of the relevant ionizable portion, where pKa is the negative logarithm of the acid dissociation constant (Ka) of a particular proton from the conjugate base.

[0060] For example, acidic residues (such as aspartic acid and glutamic acid) have side-chain carboxylic acid groups with a pKa value of approximately 4.1 (the exact pKa value can vary depending on temperature, ionic strength, and the microenvironment of the ionizable group). Therefore, these side chains exhibit a negative charge at a pH of 7.4 (often referred to as "physiological pH"). At lower pH values, these side chains protonate and lose their charge.

[0061] Conversely, basic residues such as lysine and arginine have nitrogen-containing side chains with pKa values ​​of approximately 10⁻¹². Therefore, these side chains exhibit a positive charge at pH 7.4. At higher pH values, these side chains will deprotonate and lose their charge.

[0062] Therefore, the total (net) charge of a protein molecule depends on the number of acidic and basic residues present in the protein (and their surface exposure) and the surrounding pH. Changing the surrounding pH changes the total charge on the protein. Thus, for every protein, there is a given pH at which the amount of positive and negative charge is equal, and the protein exhibits no overall net charge. This point is called the isoelectric point (pI). The isoelectric point is a standard concept in protein biochemistry familiar to those skilled in the art.

[0063] Therefore, the isoelectric point (pI) is defined as the pH at which a protein exhibits zero net charge. An increase in pI means that the protein requires a higher pH to exhibit zero net charge. Thus, an increase in pI indicates an increase in the net positive charge of the protein at a given pH. Conversely, a decrease in pI means that the protein requires a lower pH to exhibit zero net charge. Thus, a decrease in pI indicates a decrease in the net positive charge of the protein at a given pH.

[0064] Methods for determining the pI of a protein are known in the art and are familiar to those skilled in the art. For example, the pI of a protein can be calculated from the average pKa value of each amino acid present in the protein (“calculated pI”). This calculation can be performed using computer programs known in the art, such as those from ExPASy (…). https: / / web.expasy.org / compute_pi / The Compute pI / MW Tool, according to the present invention, is a preferred method for calculating pI. The same calculation technique / procedure should be used to compare pI values ​​between different molecules.

[0065] Where appropriate, the calculated pI (“observed pI”) of a protein can be experimentally confirmed using isoelectric focusing. This technique uses electrophoresis to separate proteins based on their pI. Isoelectric focusing is typically performed using a gel with a fixed pH gradient. When an electric field is applied, the protein migrates through the pH gradient until it reaches a pH at which its net charge is zero; this point is the protein's pI. The results provided by isoelectric focusing are generally of relatively low resolution by nature, therefore the inventors of this invention have found that results provided by calculated pI (as described above) are more suitable for use.

[0066] Throughout this specification, unless otherwise stated, "pI" means "calculated pI".

[0067] The pI of a protein can be increased or decreased by altering the number of basic and / or acidic groups displayed on its surface. This can be achieved by modifying one or more amino acids of the protein. For example, an increase in pI can be provided by reducing the number of acidic residues or by increasing the number of basic residues.

[0068] The modified BoNT / A or its fragments of the present invention may have a pI value that is at least 0.2, 0.4, 0.5, or 1 pI unit higher than that of unmodified BoNT / A (e.g., SEQ ID NO: 62) or its fragments. Preferably, the modified BoNT / A or its fragments may have a pI of at least 6.6, for example, at least 6.8.

[0069] The following table lists the properties of 20 standard amino acids:

[0070] amino acids side chain Aspartic acid Asp D Charged (acidic) glutamic acid Glu E Charged (acidic) Arginine Arg R Charged (alkaline) Lysine Lys K Charged (alkaline) Histidine His H Uncharged (polarity) Asparagine Asn N Uncharged (polarity) glutamine Gln Q Uncharged (polarity) Serine Ser S Uncharged (polarity) threonine Thr T Uncharged (polarity) Tyrosine Tyr Y Uncharged (polarity) Methionine Met M Uncharged (polarity) Tryptophan Trp W Uncharged (polarity) Cysteine Cys C Uncharged (polarity) alanine Ala A Non-electrostatic (hydrophobic) glycine Gly G Non-electrostatic (hydrophobic) Valine Val V Non-electrostatic (hydrophobic) Leucine Leu L Non-electrostatic (hydrophobic) Isoleucine Ile I Non-electrostatic (hydrophobic) proline Pro P Non-electrostatic (hydrophobic) Phenylalanine Phe F Non-electrostatic (hydrophobic)

[0071] The following amino acids are considered to be charged amino acids: aspartic acid (negative), glutamic acid (negative), arginine (positive), and lysine (positive).

[0072] At pH 7.4, the side chains of aspartic acid (pKa 3.1) and glutamic acid (pKa 4.1) carry a negative charge, while the side chains of arginine (pKa 12.5) and lysine (pKa 10.8) carry a positive charge. Aspartic acid and glutamic acid are referred to as acidic amino acid residues. Arginine and lysine are referred to as basic amino acid residues.

[0073] The following amino acids are considered to be uncharged polar amino acids (meaning they can participate in hydrogen bonding): asparagine, glutamine, histidine, serine, threonine, tyrosine, cysteine, methionine, and tryptophan.

[0074] The following amino acids are considered to be uncharged hydrophobic amino acids: alanine, valine, leucine, isoleucine, phenylalanine, proline, and glycine.

[0075] In amino acid insertion, additional amino acid residues (which are not normally present) are integrated into the BoNT / A polypeptide sequence or a fragment thereof, thereby increasing the total number of amino acid residues in the sequence. In amino acid deletion, amino acid residues are removed from the clostoxin amino acid sequence, thereby reducing the total number of amino acid residues in the sequence.

[0076] Preferably, the modification is a substitution, which advantageously maintains the same number of amino acid residues in the modified BoNT / A or a fragment thereof. In amino acid substitution, the amino acid residues forming a portion of the BoNT / A polypeptide sequence or a fragment thereof are replaced by different amino acid residues. As described above, the substituted amino acid residues can be one of 20 standard amino acids. Alternatively, the substituted amino acid in amino acid substitution can be a non-standard amino acid (an amino acid that is not part of the aforementioned 20 standard groups). For example, the substituted amino acid can be a basic non-standard amino acid, such as L-ornithine, L-2-amino-3-guanidinopropionic acid, or the D-isomers of lysine, arginine, and ornithine. Methods for introducing non-standard amino acids into proteins are known in the art and include recombinant protein synthesis using an *E. coli* auxotrophic expression host.

[0077] In one embodiment, the substitution is selected from: acidic amino acid residues being substituted with basic amino acid residues, acidic amino acid residues being substituted with uncharged amino acid residues, and uncharged amino acid residues being substituted with basic amino amino acid residues. In one embodiment, the substitution is wherein the acidic amino acid residue is substituted with an uncharged amino acid residue, or the acidic amino acid residue is substituted with its corresponding uncharged amide amino acid residue (i.e., aspartic acid is substituted with asparagine, and glutamic acid is substituted with glutamine).

[0078] Preferably, the basic amino acid residue is a lysine residue or an arginine residue. In other words, the substitution is with lysine or arginine. Most preferably, the modification is with lysine substitution.

[0079] Preferably, the modified BoNT / A or its fragment used in this invention contains a component located at clostoxin H. CN The modified BoNT / A or its fragments preferably contain 4 to 40 amino acid modifications. The modified BoNT / A or its fragments preferably also have a pI of at least 6.6. The modified BoNT / A preferably comprises modifications of at least four amino acids selected from the following: ASN 886, ASN 930, ASN 954, SER 955, GLN991, ASN 1025, ASN 1026, and ASN 1052, wherein the modifications include substitution with amino acids containing lysine or arginine residues. For example, the modified BoNT / A or its fragments may comprise modifications of at least five amino acids selected from the following: ASN 886, ASN 930, ASN 954, SER 955, GLN 991, ASN 1025, ASN 1026, ASN 1052, and GLN 1229, wherein the modifications include substitution with amino acids containing lysine or arginine residues.

[0080] Methods for modifying proteins by substituting, inserting, or deleting amino acid residues are known in the art. For example, amino acid modifications can be introduced by modifying the DNA sequence encoding a polypeptide (e.g., encoding unmodified BoNT / A or fragments thereof). This can be achieved using standard molecular cloning techniques, such as site-directed mutagenesis, where a polymerase is used to replace the original coding sequence with a short-chain DNA (oligonucleotide) encoding the desired amino acid, or by inserting / deleting portions of the gene with various enzymes (e.g., ligases and restriction endonucleases). Alternatively, the modified gene sequence can be chemically synthesized.

[0081] In one aspect, the present invention provides a polypeptide for promoting neuronal growth or neuronal repair to treat a subject’s neurological disorder, wherein the polypeptide comprises a polypeptide sequence having at least 70% sequence identity with SEQ ID NO:42 and / or wherein the polypeptide comprises a polypeptide sequence encoded by a nucleotide sequence having at least 70% sequence identity with SEQ ID NO:41.

[0082] In related aspects, a method for promoting neuronal growth or neuronal repair to treat a subject's neurological disorder is provided, the method comprising administering a polypeptide to the subject, wherein the polypeptide comprises a polypeptide sequence having at least 70% sequence identity with SEQ ID NO:42 and / or wherein the polypeptide comprises a polypeptide sequence encoded by a nucleotide sequence having at least 70% sequence identity with SEQ ID NO:41.

[0083] In another related aspect, the use of a polypeptide in the preparation of a medicament for promoting neuronal growth or neuronal repair to treat a subject’s neurological disorder is provided, wherein the polypeptide comprises a polypeptide sequence having at least 70% sequence identity with SEQ ID NO:42 and / or wherein the polypeptide comprises a polypeptide sequence encoded by a nucleotide sequence having at least 70% sequence identity with SEQ ID NO:41.

[0084] In one aspect, the present invention provides a polypeptide for treating a subject’s neurological disorder, wherein the polypeptide comprises a polypeptide sequence having at least 70% sequence identity with SEQ ID NO:42 and / or wherein the polypeptide comprises a polypeptide sequence encoded by a nucleotide sequence having at least 70% sequence identity with SEQ ID NO:41.

[0085] In a related aspect, a method for treating a neurological disorder in a subject is provided, the method comprising administering a polypeptide to the subject, wherein the polypeptide comprises a polypeptide sequence having at least 70% sequence identity with SEQ ID NO:42 and / or wherein the polypeptide comprises a polypeptide sequence encoded by a nucleotide sequence having at least 70% sequence identity with SEQ ID NO:41.

[0086] In another related aspect, the use of a polypeptide in the preparation of a medicament for treating a neurological disorder in a subject is provided, wherein the polypeptide comprises a polypeptide sequence having at least 70% sequence identity with SEQ ID NO:42 and / or wherein the polypeptide comprises a polypeptide sequence encoded by a nucleotide sequence having at least 70% sequence identity with SEQ ID NO:41.

[0087] In one embodiment, the polypeptide used according to the invention comprises a polypeptide sequence having at least 80%, 90%, 95%, or 98% sequence identity with SEQ ID NO:42. Preferably, the polypeptide used according to the invention comprises a polypeptide sequence as shown in SEQ ID NO:42.

[0088] In one embodiment, the polypeptide used according to the invention comprises a polypeptide sequence encoded by a nucleotide sequence having at least 80%, 90%, 95%, or 98% sequence identity with SEQ ID NO: 41. Preferably, the polypeptide used according to the invention comprises a polypeptide sequence encoded by a nucleotide sequence as shown in SEQ ID NO: 41.

[0089] In one embodiment, the polypeptide used according to the invention (e.g., comprising SEQ ID NO:42 or encoded by SEQ ID NO:41) may be a portion of a polypeptide having at least 70% sequence identity with SEQ ID NO:61 or 65. Therefore, in one embodiment, the polypeptide used according to the invention may comprise a polypeptide sequence having at least 80%, 90%, 95%, or 98% sequence identity with SEQ ID NO:61 or 65. Preferably, the polypeptide used according to the invention may comprise SEQ ID NO:61 or 65 (more preferably composed of them). In one embodiment, the polypeptide comprises a non-catalytically active L-chain (e.g., according to SEQ ID NO:65).

[0090] In one embodiment, the polypeptide used according to the invention (e.g., comprising SEQ ID NO:42 or encoded by SEQ ID NO:41) can be encoded by a nucleotide sequence having at least 70% sequence identity with SEQ ID NO:60. Therefore, in one embodiment, the polypeptide used according to the invention can be encoded by a nucleotide sequence having at least 80%, 90%, 95%, or 98% sequence identity with SEQ ID NO:60. Preferably, the polypeptide used according to the invention can be encoded by a nucleotide sequence comprising SEQ ID NO:60 (more preferably composed of thereof). In one embodiment, the polypeptide comprises a non-catalytically active L-chain.

[0091] SEQ ID NO:42 is an example of a modified BoNT / A fragment, while SEQ ID NO:61 and 65 are examples of modified BoNT / A peptides with and without catalytic activity, respectively. Such modified BoNT / A peptides and fragments are particularly preferred for use in this invention. Compared to wild-type BoNT / A, the peptides shown in SEQ ID NO:42, 61, and 62 have numerous amino acid modifications (e.g., substitutions), which increase the isoelectric point of the peptides. Without being bound by theory, it is assumed that the increased net positive charge promotes electrostatic interactions between the peptide and the anionic extracellular component, thereby promoting binding between the peptide and the cell surface, thus increasing retention and / or duration of action at the site of application. Therefore, it is envisioned that the neuronal growth and / or repair properties of SEQ ID NO:42, 61, and 65 will be enhanced compared to equivalent peptides lacking the aforementioned modifications.

[0092] For the aforementioned catalytically modified BoNT / A peptides (e.g., SEQ ID NO: 61), one way to define these advantageous properties (which represent an increase in the therapeutic index) is in terms of the safety ratio of the modified BoNT / A. In this regard, the adverse effects of clostridial neurotoxins (caused by toxin diffusion from the application site) can be experimentally assessed by measuring the percentage of weight loss in relevant animal models (e.g., mice, where weight loss is detected within seven days of administration). Conversely, the desired targeting effect of the clostridial neurotoxin can be experimentally assessed by a digital abduction score (DAS) assay (a measure of muscle paralysis). The DAS assay can be performed by injecting 20 μl of clostridial neurotoxin prepared in gelatin phosphate buffer into the gastrocnemius / soleus muscle complex of mice, followed by assessment of the digital abduction score using Aoki's method (Aoki KR, Toxicon 39:1815-1820; 2001). In the DAS assay, mice were briefly suspended by their tails to elicit a characteristic startle response in which they extended their hind limbs and abducted their hind fingers. The degree of finger abduction following clostridial neurotoxin injection was scored on a five-point scale (0 = normal to 4 = maximum reduction in finger abduction and leg extension).

[0093] The safety ratio of clostridial neurotoxin can then be expressed as the ratio between the amount of toxin required for a 10% weight loss (measured within the first seven days after administration to mice when peak effect is achieved) and the amount of toxin required for a DAS score of 2. Therefore, a high safety ratio score is required, indicating that the toxin effectively paralyzes the target muscle with minimal undesirable off-target effects. The catalytically modified BoNT / A of this invention can have a higher safety ratio than equivalent unmodified (natural) botulinum toxins (e.g., SEQ ID NO: 62).

[0094] Therefore, in one embodiment, the catalytically modified BoNT / A of the present invention has a safety ratio of at least 8 (e.g., at least 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, or 50), wherein the safety ratio is calculated as: the toxin dose (pg / mouse) required for -10% body weight change divided by DAS ED. 50 (pg / mouse) [ED50 = the dose required to produce a DAS score of 2].

[0095] In one embodiment, the catalytically modified BoNT / A of the present invention has a safety ratio of at least 10. In another embodiment, the modified BoNT / A or a fragment thereof of the present invention has a safety ratio of at least 15.

[0096] A polypeptide containing at least 70% sequence identity with SEQ ID NO:61 is described in WO 2015 / 004461A1, which is incorporated herein by reference in its entirety.

[0097] In one embodiment, a polypeptide comprising a polypeptide sequence having at least 70% sequence identity with SEQ ID NO:42, 61, or 65 and / or comprising a polypeptide sequence encoded by a nucleotide sequence having at least 70% sequence identity with SEQ ID NO:41 or 60 includes substitutions at one or more (preferably two or more, three or more, four or more, five or more, or six or more, more preferably all) positions of 930, 955, 991, 1026, 1052, 1229, and 886. The position number corresponds to the position of SEQ ID NO:62 and can be determined by comparing the polypeptide sequence with SEQ ID NO:62 (unmodified / wild-type BoNT / A). Since the presence of a methionine residue at position 1 of SEQ ID NO:62 is optional, a person skilled in the art will consider the presence / absence of the methionine residue when determining the amino acid residue number. For example, when SEQ ID NO:62 includes methionine, the position number will be as defined above (e.g., position 886 would be ASN 886 of SEQ ID NO:62). Alternatively, if methionine is absent in SEQ ID NO:62, the amino acid residue number should be changed by -1 (e.g., position 886 would be ASN885 of SEQ ID NO:62). Similar considerations apply to the presence / absence of methionine at position 1 in other polypeptide sequences described herein, and those skilled in the art will readily determine the correct amino acid residue number using conventional techniques in the art.

[0098] Preferably, the polypeptide comprising a polypeptide sequence having at least 70% sequence identity with SEQ ID NO:42, 61, or 65 and / or comprising a polypeptide sequence encoded by a nucleotide sequence having at least 70% sequence identity with SEQ ID NO:41 or 60 contains lysine or arginine (more preferably lysine) at one or more of positions 930, 955, 991, 1026, 1052, 1229, and 886. In one embodiment, the polypeptide contains lysine or arginine (more preferably lysine) at at least two, three, four, five, six, or all of positions 930, 955, 991, 1026, 1052, 1229, and 886. Most preferably, the polypeptide contains lysine or arginine (more preferably lysine) at all positions 930, 955, 991, 1026, 1052, 1229, and 886.

[0099] The polypeptides of this invention promote neuronal growth and / or neuronal repair. Therefore, the polypeptides can be used to treat neurological disorders. As used herein, the term "neurological disorder" is a disorder that can be treated by promoting neuronal growth and / or repair in a subject.

[0100] Therefore, in one aspect, the present invention provides a method for promoting neuronal growth and / or neuronal repair, the method comprising administering a polypeptide to a subject, the polypeptide comprising a clostridium neurotoxin light chain (L-chain) or a fragment thereof; and / or a fragment of a clostridium neurotoxin heavy chain (H-chain). In another aspect, the present invention provides a method for promoting neuronal growth and / or neuronal repair, the method comprising administering a polypeptide to a subject, the polypeptide comprising a catalytically inactive clostridium neurotoxin L-chain. In yet another aspect, a method for promoting neuronal growth or neuronal repair is provided, the method comprising administering a polypeptide to a subject, wherein the polypeptide comprises a polypeptide sequence having at least 70% sequence identity with SEQ ID NO:42 and / or wherein the polypeptide comprises a polypeptide sequence encoded by a nucleotide sequence having at least 70% sequence identity with SEQ ID NO:41. In yet another aspect, a method for promoting neuronal growth or neuronal repair is provided, the method comprising administering a polypeptide to a subject, wherein the polypeptide comprises a polypeptide sequence having at least 70% sequence identity with SEQ ID NO:63.

[0101] The term "promote neuronal growth and / or neuronal repair" may mean that the peptides of the present invention initiate neuronal growth and / or neuronal repair, for example, in the absence of neuronal growth and / or neuronal repair. In other embodiments, the term "promote neuronal growth and / or neuronal repair" may mean that the peptides of the present invention increase the rate of neuronal growth and / or neuronal repair. This increase may be an increase compared to the rate of neuronal growth and / or neuronal repair in the absence of the peptides of the present invention. In one embodiment, neuronal growth and / or neuronal repair allows for the reconstruction of damaged neuronal circuits, thereby restoring activity and / or neuronal communication in a neuronal network or group of neurons. Therefore, the term "neural repair" as used herein may encompass both the repair of a specific neuron and the repair of a neuronal circuit.

[0102] The term "neuronal growth and / or neuronal repair" may also include neuronal plasticity. Therefore, in one embodiment, the peptides of the present invention promote neuronal plasticity. As used herein, the term "neuronal plasticity" includes axonal budding, dendritic budding, neurogenesis (e.g., the generation of new neurons), maturation, differentiation, and / or synaptic plasticity (e.g., including changes in synaptic strength, activity, anatomy, and / or connectivity). In one embodiment, the peptides of the present invention promote the establishment of functional synapses (e.g., at or near the site of injury).

[0103] Compared to neuronal growth and / or repair in the absence of the polypeptide of the present invention or in the presence of an alternative polypeptide, neuronal growth and / or repair can be increased by at least 10%, 20%, 30%, 40%, 50%, 60%, or 70% (preferably at least 80%) in the presence of the polypeptide of the present invention. In some embodiments, compared to neuronal growth and / or repair in the absence of the polypeptide of the present invention or in the presence of an alternative polypeptide, neuronal growth and / or repair can be increased by at least 100%, 150%, or 200% in the presence of the polypeptide of the present invention.

[0104] In one embodiment, the polypeptides of the present invention promote neuronal growth. As used herein, the term "neuronal growth" encompasses the growth of any part of a neuron, including the growth of axons and / or dendrites. The polypeptides of the present invention can increase neurite length, neurite number (e.g., the number of neurites per cell), and / or can increase the length and / or number of processes originating from the cell body or cell membrane of the neuron. Preferably, the polypeptides of the present invention promote axonal growth of neurons (e.g., neurons of a subject). In other words, the polypeptides of the present invention preferably increase axonal growth, such as increasing axonal budding. This axonal growth can promote connections and / or chemical communication between neurons.

[0105] The neurological disorders treated by the peptides of the present invention can be neuronal damage, neurodegenerative disorders, sensory disorders, or autonomic disorders.

[0106] Neurological disorders can be neuronal damage. In one implementation, neuronal damage can be nerve injury, neuropathy (e.g., peripheral neuropathy), spinal cord injury, nerve transection, brain injury (e.g., traumatic brain injury), non-traumatic injury (e.g., stroke or spinal cord infarction), or brachial plexus injury, such as Erb paralysis or Klumpke paralysis.

[0107] In one embodiment, nerve trauma may be caused by scarring and / or fracture. In such cases of nerve trauma, nerve endings are damaged. The polypeptides of the present invention advantageously allow for the repair of said nerve endings or allow for the repair of distal nerve endings that allow for the treatment of nerve damage.

[0108] Neuronal damage can cause paralysis, such as paralysis caused by spinal cord injury (e.g., caused by compression, contraction, and / or stretching). In one implementation, spinal cord injury is paraplegia or quadriplegia.

[0109] Neurological disorders can be sensory disorders. In one embodiment, the sensory disorder is sensory neuropathy, sensorimotor polyneuropathy, diabetic neuropathy, pain, Brown-Sequard syndrome, Charcot-Marie-Tooth disease, or Devic syndrome. Preferably, the sensory disorder described herein is not pain. In other words, the neurological disorder described herein is preferably not pain.

[0110] Neurological disorders can be autonomic disorders. In one implementation, an autonomic disorder is autonomic neuropathy, multiple system atrophy, acute idiopathic polyneuropathy, autonomic dysfunction, familial autonomic dysfunction, diabetic autonomic failure, isolated autonomic failure, thermoregulation disorder, hyperhidrosis, neurally mediated syncope (vasovagal, urination, coughing, swallowing, and other situational forms), erectile dysfunction, orthostatic hypotension, postural tachycardia syndrome (PoTS), or Guillain-Barre syndrome.

[0111] Neurological disorders can be neurodegenerative disorders. In one embodiment, the neurodegenerative disorder is Alzheimer's disease, Parkinson's disease, Parkinson's disease-related disorders, motor neuron disease, peripheral neuropathy, motor neuropathy, prions, Huntington's disease, spinocerebellar ataxia, spinal muscular atrophy, unilateral muscular atrophy, Friedreich's ataxia, Hallervorden-Spatz disease, or frontotemporal degeneration. Preferably, the neurodegenerative disorder is Parkinson's disease or motor neuron disease. Advantageously, the peptides of the present invention are considered useful for treating neurodegenerative disorders, thereby restoring activity and / or neuronal communication in neuronal networks or groups of neurons, due to their ability to promote neuronal growth (e.g., including neuronal plasticity) and / or neuronal repair, and further due to their ability to reconstruct damaged neuronal circuits.

[0112] Given its ability to promote neuronal growth and / or neuronal repair, the polypeptides of the present invention can be considered neurotrophic polypeptides. The neurons described herein can be selected from one or more of the following: motor neurons (including autonomic neurons), sensory neurons, spinal interneurons, and brain interneurons. Therefore, in one embodiment, the polypeptides of the present invention promote the growth and / or repair of motor neurons, sensory neurons, and / or interneurons. Preferably, the polypeptides of the present invention promote the growth and / or repair of motor neurons.

[0113] As used herein, “subject” can be a mammal, such as a human or other mammal. Preferably, “subject” refers to a human subject.

[0114] As used herein, the term "barrier" also includes "disease." In one implementation, the barrier is a disease.

[0115] As used herein, the term "treat" or "treating" includes preventative treatment (e.g., preventing the onset of a disorder) and corrective treatment (treating a subject who already has a disorder). Preferably, as used herein, "treat" or "treating" refers to corrective treatment.

[0116] As used in this article, the term “treat” or “treating” refers to an obstacle and / or its symptoms.

[0117] Therefore, the polypeptide of the present invention can be administered to subjects at a therapeutically effective amount or a preventatively effective amount. Preferably, the polypeptide of the present invention is administered to subjects at a therapeutically effective amount.

[0118] "Therapeutic effective amount" is any amount of a polypeptide, whether administered alone or in combination, sufficient to achieve such treatment of the disorder (or its symptoms) when administered to a subject to treat the disorder (or its symptoms).

[0119] "Preventive effective dose" is any amount of a polypeptide, administered alone or in combination to a subject to suppress or delay the onset or recurrence of a disorder (or its symptoms). In some implementations, the preventive effective dose completely prevents the onset or recurrence of the disorder. "Suppressing" an onset means reducing the likelihood of an onset of the disorder (or its symptoms) or completely preventing an onset of the disorder.

[0120] The polypeptides of the present invention can be formulated in any suitable manner for administration to a subject, for example, as part of a pharmaceutical composition. Thus, in one aspect, the present invention provides a pharmaceutical composition comprising the polypeptides of the present invention and a pharmaceutically acceptable carrier, excipient, adjuvant, propellant, and / or salt. In some embodiments, the polypeptides of the present invention may be in a single-chain form, while in other embodiments, the polypeptides may be in a double-chain form, for example, wherein the two chains are connected by a disulfide bridge. Preferably, the polypeptide is in a double-chain form.

[0121] The polypeptides of the present invention can be formulated for oral, parenteral, continuous infusion, inhalation, or topical application. Compositions suitable for injection can be in the form of solutions, suspensions, or emulsions, or in the form of dry powder dissolved or suspended in a suitable carrier prior to use.

[0122] In the case of locally delivered peptides, the peptides can be formulated into creams (e.g. for topical application) or for subcutaneous injection.

[0123] Local delivery methods may include aerosols or other sprays (e.g., nebulizers). In this regard, aerosol formulations of peptides can be delivered to the lungs and / or other nasal and / or bronchial or airway pathways.

[0124] The polypeptides of the present invention can be administered to subjects via intrathecal or epidural injection into the spinal segment at the level of the spinal segment involving the nerve innervation of the affected organ.

[0125] The route of administration can be via laparoscopy and / or local injection. In one embodiment, the polypeptide of the present invention is administered at or near the site of injury, preferably at the site of injury. For example, in the case of a spinal cord injury, the polypeptide can be administered intrathecally or intraspinally (preferably intrathecally). In one embodiment, the route of administration of the polypeptide of the present invention can be perineural, intraneural, intraspinal, and / or intrathecal.

[0126] The dosage range of the polypeptides of this invention is the dosage range that produces the desired therapeutic and / or preventive effects. It should be understood that the required dosage range depends on the exact nature of the clostridium neurotoxin or composition, the route of administration, the nature of the formulation, the age of the subject, the nature, degree, or severity of the subject's condition, contraindications, if any, and the judgment of the attending physician. Variations in these dosage levels can be adjusted for optimization using standard empirical procedures.

[0127] In one embodiment, the dose of the peptide is a fixed dose. The fixed dose can be in the range of 50 pg to 250 μg, preferably 100 pg to 100 μg. In one embodiment, the fixed dose can be at least 50 pg, 100 pg, 500 pg, 1 ng, 50 ng, 100 ng, 500 ng, 1 μg, or 50 μg. The dose can be a single fixed dose.

[0128] Fluid dosage forms are typically prepared using peptides and pyrogen-free sterile carriers. Depending on the carrier and concentration used, clostridial neurotoxins can be dissolved or suspended in the carrier. In preparing the solution, the peptide can be dissolved in the carrier, and if necessary, the solution can be made isotonic by adding sodium chloride, and sterilized using aseptic techniques by filtration through a sterile filter before being filled into suitable sterile vials or ampoules and sealed. Alternatively, if the solution is sufficiently stable, the solution in the sealed container can be sterilized by autoclaving. Advantageously, additives such as buffers, solubilizers, stabilizers, preservatives or bactericides, suspending agents or emulsifiers, and / or local anesthetics can be dissolved in the carrier.

[0129] Dry powder dissolved or suspended in a suitable carrier before use can be prepared by filling pre-sterilized ingredients into sterile containers using aseptic techniques in a sterile area. Alternatively, the ingredients can be dissolved into suitable containers using aseptic techniques in a sterile area. The product is then freeze-dried and the containers are aseptically sealed.

[0130] The parenteral suspensions suitable for the routes of administration described herein are prepared in substantially the same manner, except that the sterile components are suspended in a sterile carrier rather than dissolved and sterilization is not achieved by filtration. The components may be separated under sterile conditions, or alternatively, sterilized after separation, for example, by gamma irradiation.

[0131] Advantageously, a suspending agent (e.g., polyvinylpyrrolidone) is included in the composition to promote uniform distribution of the components.

[0132] The drug delivery according to the present invention can utilize a variety of delivery technologies, including microparticle encapsulation or high-pressure aerosol impingement.

[0133] The polypeptides of the present invention may be clostridium neurotoxin or fragments thereof, preferably fragments thereof.

[0134] In one embodiment, the polypeptide of the present invention may be encoded by a nucleotide sequence having at least 70% sequence identity with any one of SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49 or 60. In one embodiment, the polypeptide of the present invention may be encoded by a nucleotide sequence having at least 80%, 90%, 95% or 98% sequence identity with any one of SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49 or 60. Preferably, the polypeptide of the present invention may be encoded by a nucleotide sequence comprising any one of SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49 or 60.

[0135] In one embodiment, the polypeptide of the present invention may comprise a polypeptide sequence having at least 70% sequence identity with any one of SEQ ID NO:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 61, 62, 63, 64 or 65. In one embodiment, the polypeptide of the present invention may comprise a polypeptide sequence having at least 80%, 90%, 95%, or 98% sequence identity with any one of SEQ ID NO:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 61, 62, 63, 64, or 65. Preferably, the polypeptide of the present invention may comprise a polypeptide sequence of any one of SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 61, 62, 63, 64 or 65.

[0136] In one embodiment, the invention includes the use of a full-length clostridial neurotoxin comprising a clostridial neurotoxin L-chain and a clostridial neurotoxin H-chain, provided that the clostridial neurotoxin L-chain is non-catalytically active.

[0137] The term "clostridium neurotoxin" includes toxins produced by Clostridium botulinum (botulinum neurotoxin serotypes A, B, C1, D, E, F, G, and X), Clostridium tetani (tetanus neurotoxin), Clostridium butyricum (botulinum neurotoxin serotype E), and Clostridium baratii (botulinum neurotoxin serotype F), as well as modified clostridium neurotoxins or derivatives derived from any of the above.

[0138] Botulinum neurotoxin (BoNT) is produced by Clostridium botulinum as a large protein complex, consisting of the BoNT itself in combination with numerous accessory proteins. Currently, there are eight different classes of botulinum neurotoxin: serotypes A, B, C1, D, E, F, G, and X, all with similar structures and modes of action. Different BoNT serotypes can be distinguished by inactivation by specific neutralizing antisera; this serotype classification is related to the percentage of sequence identity at the amino acid level. Based on this percentage of sequence identity, a given serotype of BoNT protein is further subdivided into different subtypes.

[0139] BoNT is absorbed in the gastrointestinal tract and, upon entering systemic circulation, binds to the presynaptic membrane of cholinergic nerve endings, preventing the release of the neurotransmitter acetylcholine. BoNT / B, BoNT / D, BoNT / F, and BoNT / G cleave small synaptic vesicle proteins / vesicle-associated membrane proteins (VAMPs); BoNT / C1, BoNT / A, and BoNT / E cleave 25 kDa (SNAP-25) synaptosome-associated proteins; BoNT / C1 cleaves synaptic proteins. BoNT / X has been found to cleave SNAP-25, VAMP1, VAMP2, VAMP3, VAMP4, VAMP5, Ykt6, and syntaxin1.

[0140] Tetanus toxin is produced by Clostridium tetani in a single serotype. Clostridium butyricum produces BoNT / E, while Clostridium balana produces BoNT / F.

[0141] The term "clostridium neurotoxin" is also intended to include modified clostridium neurotoxins and their derivatives, including but not limited to those described below. Modified clostridium neurotoxins or derivatives may contain one or more amino acids that have been modified compared to the natural (unmodified) form of clostridium neurotoxin, or may contain one or more inserted amino acids not present in the natural (unmodified) form of clostridium neurotoxin. For example, a modified clostridium neurotoxin may have a modified amino acid sequence in one or more domains relative to the sequence of a natural (unmodified) clostridium neurotoxin. Such modification can alter functional aspects of the toxin, such as biological activity or persistence. Therefore, in one embodiment, the clostridium neurotoxin of the present invention is a modified clostridium neurotoxin, or a modified clostridium neurotoxin derivative, or a clostridium neurotoxin derivative.

[0142] Modified clostridium neurotoxins can have one or more modifications in the amino acid sequence of the heavy chain (such as modified H). C (structural domain), wherein the modified heavy chain binds to target nerve cells with a higher or lower affinity than natural (unmodified) clostridium neurotoxin. H C Such modifications in a structural domain can include modifications to H C Residues in the ganglioside-binding site or protein (SV2 or synaptic binding protein) binding site of the domain, which are altered to bind to ganglioside receptors and / or protein receptors on target nerve cells. Examples of such modified clostridial neurotoxins are described in WO 2006 / 027207 and WO2006 / 114308, both of which are incorporated herein by reference in their entirety.

[0143] Modified clostridial neurotoxins may have one or more modifications in the amino acid sequence of the light chain, such as modifications in the substrate-binding or catalytic domains, which can alter or modify the SNARE protein specificity of the modified L-chain. Examples of such modified clostridial neurotoxins are described in WO 2010 / 120766 and US2011 / 0318385, both of which are incorporated herein by reference in their entirety.

[0144] Modified clostridial neurotoxins may contain one or more modifications that increase or decrease the biological activity and / or biological persistence of the modified clostridial neurotoxin. For example, modified clostridial neurotoxins may contain leucine- or tyrosine-based motifs that increase or decrease the biological activity and / or biological persistence of the modified clostridial neurotoxin. Suitable leucine-based motifs include xDxxxLL, xExxxLL, xExxxIL, and xExxxLM (where x is any amino acid). Suitable tyrosine-based motifs include Yxx-Hy (where Hy is a hydrophobic amino acid). Examples of modified clostridial neurotoxins containing leucine- and tyrosine-based motifs are described in WO 2002 / 08268, which is incorporated herein by reference in its entirety.

[0145] As described above, a modified clostridial neurotoxin (or clostridial neurotoxin fragment) can be a toxin containing one or more modifications, the modifications of which increase the isoelectric point of the clostridial neurotoxin compared to an equivalent unmodified clostridial neurotoxin lacking the one or more modifications. Suitable modified clostridial neurotoxins are described above and in WO 2015 / 004461 A1 and WO2016 / 110662 A1, which are incorporated herein by reference. Exemplary sequences include SEQ ID NO: 61 and 42 as described herein.

[0146] The term "clostridium neurotoxin" is intended to include both heterozygous and chimeric clostridium neurotoxins. A heterozygous clostridium neurotoxin comprises at least a portion of a light chain from one clostridium neurotoxin or its subtype, and at least a portion of a heavy chain from another clostridium neurotoxin or its subtype. In one embodiment, a heterozygous clostridium neurotoxin may contain a complete light chain from one clostridium neurotoxin subtype and a heavy chain from another clostridium neurotoxin subtype. In another embodiment, a chimeric clostridium neurotoxin may contain a portion of a heavy chain (e.g., a binding domain) from one clostridium neurotoxin subtype, while another portion of the heavy chain originates from another clostridium neurotoxin subtype. Similarly or alternatively, a therapeutic element may comprise light chain portions from different clostridium neurotoxins. Such hybrid or chimeric clostridial neurotoxins are useful, for example, as a method of delivering the therapeutic benefits of these clostridial neurotoxins to subjects who are immune to a given clostridial neurotoxin subtype, subjects who may have below-average receptor concentrations to a given clostridial neurotoxin heavy chain binding domain, or subjects who may have protease-resistant variants of membrane or vesicular toxin substrates (e.g., SNAP-25, VAMP, and synaptic proteins). Hybrid and chimeric clostridial neurotoxins are described in US 8,071,110, which is incorporated herein by reference in its entirety. Thus, in one embodiment, the clostridial neurotoxin (or fragment thereof) of the present invention is a hybrid or chimeric clostridial neurotoxin.

[0147] In a particularly preferred embodiment, the polypeptide of the present invention may be a chimeric clostridium neurotoxin comprising (preferably consisting of) a BoNT / A light chain and a translocation domain, and a BoNT / B receptor-binding domain (HC domain) or a portion thereof. Suitable chimeric and / or hybrid clostridium neurotoxins may be one of those taught in WO 2017 / 191315A1, which is incorporated herein by reference. Such preferred sequences include SEQ ID NO:44, 63, and 64.

[0148] BoNT / A LH N Domains can be covalently connected to BoNT / BH C Structural domain. The chimera BoNT / A is also referred to herein as “BoNT / AB” or “BoNT / AB chimera”.

[0149] LH N The C-terminal amino acid residue of the domain can correspond to the LH that separates BoNT / A. N and HC domain 3 10 The first amino acid residue of the helix, and H C The N-terminal amino acid residue of the domain can correspond to the LH that separates BoNT / B. N and H C 3 of the structural domain 10 The second amino acid residue of the helix.

[0150] The "LH separating BoNT / A" mentioned in this article N and HC domain 3 10 The first amino acid residue of the helix refers to the LH separator. N and H C 3 of the structural domain 10 The N-terminal residue of the helix.

[0151] The "LH separating BoNT / B" mentioned in this article N and HC domain 3 10 The second amino acid residue of the helix refers to the LH separator. N and H C 3 of the structural domain 10 The amino acid residue following the N-terminal residue of the helix.

[0152] 3 10 A "helix" is a secondary structure found in proteins and polypeptides, along with α-helices, β-sheets, and inversions. 3 10The amino acids in the helix are arranged in a right-handed helical structure, where each complete turn is accomplished by three residues and ten atoms, which separate the intramolecular hydrogen bonds between them. Each amino acid corresponds to a 120° turn in the helix (i.e., three residues per turn), translating along the helical axis. (=0.2nm), and has 10 atoms in the ring formed by hydrogen bonding. Most importantly, the NH group of the amino acid forms hydrogen bonds with the C=O groups of the amino acids three residues prior; this repeating i+3→i hydrogen bond defines 3 10 Spiral. 3 10 Helices are a standard concept in structural biology that is familiar to technicians.

[0153] This 3 10 The helix corresponds to the four residues that form the actual helix and two cap (or transition) residues, with one cap at each of the four residues. As used herein, the term "separated LH" refers to... N and H C 3 of the structural domain 10 The "helix" is composed of these 6 residues.

[0154] Structural analysis and sequence alignment were performed to identify the LH separator. N and H C 3 of the structural domain 10 Spiral. This 3 10 The helix at its N-terminus (i.e., at LH) N The C-terminal portion of the structural domain is surrounded by an α-helix, and at its C-terminus (i.e., at the H... C The N-terminal portion is surrounded by a β-chain. 3 10 The first (N-terminal) residue (cap or transition residue) of the helix also corresponds to the C-terminal residue of this α-helix.

[0155] Separating LH N and H C 3 of the structural domain 10 The helix can be determined, for example, from the crystal structures of publicly available botulinum neurotoxins, such as 3BTA (http: / / www.rcsb.org / pdb / explore / explore.do?structureId=3BTA) and 1EPW (http: / / www.rcsb.org / pdb / explore / explore.do?structureId=1EPW) for botulinum neurotoxins A1 and B1, respectively.

[0156] Publicly available computer modeling and comparison tools can also be used to identify LH septa in other neurotoxins. N and H C 3 of the structural domain10 The location of helices, such as the homology modeling server LOOPP (learns, observes, and outputs protein patterns, http: / / loopp.org), PHYRE (protein homology / mimetic Y recognition engine, http: / / www.sbg.bio.ic.ac.uk / phyre2 / ) and Rosetta (https: / / www.rosettacommons.org / ), the protein stacking server SuperPose (http: / / wisart.biology.ualberta.ca / superpose / ), the alignment program Clustal Omega (http: / / www.clustal.org / omega / ), and many other tools / services listed in the Molecular and Cell Biologists' Internet Resources (http: / / molbiol-tools.ca / ). Especially "H N / H CN "The area around the junction is structurally highly conservative, making it an ideal region for superimposing different serotypes."

[0157] For example, the following methods can be used to identify this type of neurotoxin in other neurotoxins. 10 Spiral sequence:

[0158] 1. Based on the BoNT / A1 crystal structure (3BTA.pdb), the structural homology modeling tool LOOP (http: / / loopp.org) was used to obtain the predicted structures of other BoNT serotypes;

[0159] 2. Edit the resulting structure (pdb) file to include only H... CN The N-terminus of the domain and the approximately 80 residues preceding it (these are H) N (part of the structural domain), thus preserving the structurally highly conserved "H" N / H CN "area;

[0160] 3. Use the protein stacking server SuperPose (http: / / wishart.biology.ualberta.ca / superpose / ) to stack each serotype onto the 3BTA.pdb structure;

[0161] 4. Check the overlaid PDB files, and modify BoNT / A1 H. C The 3 at the beginning of the structure domain 10 Helical localization was performed, and then the corresponding residues in other serotypes were identified;

[0162] 5. Compare other BoNT serotype sequences with Clustal Omega to check if the corresponding residues are correct.

[0163] LH determined by this method N H C and 3 10 Examples of spiral structural domains are presented below:

[0164]

[0165]

[0166] Using structural analysis and sequence alignment, the LH separator was discovered. N and H C 3 of the structural domain 10 The β-chain following the helix is ​​a conserved structure in all botulinum toxins and tetanus neurotoxins, and it separates from the LH... N and H C 3 of the structural domain 10 The helix begins at the 8th residue (e.g., at residue 879 for BoNT / A1).

[0167] BoNT / AB chimeras can contain H from BoNT / B C LH from BoNT / A are covalently connected to the structural domain. N structural domain

[0168] ·Among them, LH N The C-terminal amino acid residue of the domain corresponds to the H located at BoNT / A. C The eighth amino acid residue at the N-terminus of the β-chain, which begins with the domain (N-terminus), and

[0169] ·Among them, H C The N-terminal amino acid residue of the domain corresponds to the H located in BoNT / B. C The seventh amino acid residue at the N-terminus of the β-chain, which begins with a domain (N-terminus).

[0170] BoNT / AB chimeras can contain H with BoNT / B C LH of BoNT / A with covalently connected structural domains N structural domain

[0171] ·Among them, LH N The C-terminal amino acid residue of the domain corresponds to the LH located at BoNT / A. N The C-terminal amino acid residue of the α-helix at the end of the domain (C-terminus), and

[0172] ·Among them, HC The N-terminal amino acid residue of the domain corresponds to the LH located immediately adjacent to BoNT / B. N The C-terminal amino acid residue of the C-terminal α-helix at the end of the domain (C-terminus).

[0173] The fundamental principle of the BoNT / AB chimera design process is to attempt to ensure that the secondary structure remains unaffected, thereby minimizing any changes to the tertiary structure and the function of each structural domain. Not wanting to be bound by theory, it is assumed that by not disrupting the 3... 10 The four central amino acid residues of the helix ensure the optimal conformation of the chimeric neurotoxin, thus allowing it to fully function.

[0174] LH from BoNT / A N The domain may correspond to amino acid residues 1 to 872 of SEQ ID NO:62, or a polypeptide sequence having at least 70% sequence identity with it. LH from BoNT / A N The domain may correspond to amino acid residues 1 to 872 of SEQ ID NO:62, or a polypeptide sequence having at least 80%, 90%, or 95% sequence identity with it. Preferably, LH from BoNT / A N The domains correspond to amino acid residues 1 to 872 of SEQ ID NO:62.

[0175] H from BoNT / B C The domain may correspond to amino acid residues 860 to 1291 of SEQ ID NO:52, or a polypeptide sequence having at least 70% sequence identity with it. H from BoNT / B C The domain may correspond to amino acid residues 860 to 1291 of SEQ ID NO:52, or a polypeptide sequence having at least 80%, 90%, or 95% sequence identity with it. Preferably, the H domain is derived from BoNT / B. C The domain corresponds to amino acid residues 860 to 1291 of SEQ ID NO:52.

[0176] Preferably, the BoNT / AB chimera contains BoNT / ALH. N Structural domains and BoNT / BH C Structural domain. More preferably, LH N The structural domain corresponds to amino acid residues 1 to 872 of BoNT / A (SEQ ID NO: 62), while H C The domain corresponds to amino acid residues 860 to 1291 of BoNT / B (SEQ ID NO: 52).

[0177] Preferred, BoNT / BHC Domain in H CC The subdomain also contains at least one amino acid residue substitution, addition, or deletion, which, compared to the native BoNT / B sequence, increases the binding affinity of BoNT / B neurotoxin to human Syt II. BoNT / BH CC Suitable substitutions, additions or deletions of amino acid residues in the subdomains have been disclosed in WO 2013 / 180799 and WO2016 / 154534 (both incorporated herein by reference).

[0178] BoNT / BH CC Suitable amino acid residue substitutions, additions, or deletions in the subdomains include substitution mutations selected from the following: V1118M; Y1183M; E1191M; E1191I; E1191Q; E1191T; S1199Y; S1199F; S1199L; S1201V; E1191C, E1191V, E1191L, E1191Y, S1199W, S1199E, S1199H, W1178Y, W1178Q, W1178A, W1178S, Y1183C, Y1183P, and combinations thereof.

[0179] BoNT / BH CC Suitable amino acid residue substitutions, additions, or deletions in the subdomains further include combinations of two substitution mutations selected from the following: E1191M and S1199L, E1191M and S1199Y, E1191M and S1199F, E1191Q and S1199L, E1191Q and S1199Y, E1191Q and S1199F, E1191M and S1199W, E1191M and W1178Q, E1191C and S1199W, E1191C and S1199Y, E1191C and W1178Q, E1191Q and S1199W, E1191V and S1199W, E1191V and S1191Y, or E1191V and W1178Q.

[0180] BoNT / BH CC Suitable amino acid residue substitutions, additions, or deletions in the subdomains also include combinations of three substitution mutations: E1191M, S1199W, and W1178Q.

[0181] Preferred, BoNT / BH CC Suitable amino acid residue substitutions, additions, or deletions in the subdomain include combinations of two substitution mutations, namely E1191M and S1199Y.

[0182] The modification can be a comparison with the unmodified BoNT / B as shown in SEQ ID NO:52, wherein the amino acid residue numbering is determined by comparison with SEQ ID NO:52. Since the presence of the methionine residue at position 1 of SEQ ID NO:52 is optional, those skilled in the art will consider the presence / absence of the methionine residue when determining the amino acid residue numbering. For example, in the case where SEQ ID NO:52 includes methionine, the position numbering will be as defined above (e.g., E1191 would be E1191 of SEQ ID NO:52). Alternatively, in the case where methionine is absent in SEQ ID NO:52, the amino acid residue numbering should be changed by -1 (e.g., E1191 would be E1190 of SEQ ID NO:52). Similar considerations apply to the presence / absence of methionine at position 1 in other polypeptide sequences described herein, and those skilled in the art will readily determine the correct amino acid residue numbering using techniques conventional in the art.

[0183] Therefore, in one aspect, the present invention provides a polypeptide for promoting neuronal growth or neuronal repair to treat neurological disorders in a subject, wherein the polypeptide comprises a polypeptide sequence having at least 70% sequence identity with SEQ ID NO:63 or 64.

[0184] In a related aspect, a method for promoting neuronal growth or neuronal repair to treat a neurological disorder in a subject is provided, the method comprising administering a polypeptide to the subject, wherein the polypeptide comprises a polypeptide sequence having at least 70% sequence identity with SEQ ID NO:63 or 64.

[0185] In a further related aspect, the use of the polypeptide in the preparation of a medicament for promoting neuronal growth or neuronal repair to treat neurological disorders in a subject is provided, wherein the polypeptide comprises a polypeptide sequence having at least 70% sequence identity with SEQ ID NO:63 or 64.

[0186] In one aspect, the present invention provides a polypeptide for treating a subject’s neurological disorder, wherein the polypeptide comprises a polypeptide sequence having at least 70% sequence identity with SEQ ID NO:63 or 64.

[0187] In a related aspect, a method for treating a subject’s neurological disorder is provided, the method comprising administering a polypeptide to the subject, wherein the polypeptide comprises a polypeptide sequence having at least 70% sequence identity with SEQ ID NO:63 or 64.

[0188] In another related aspect, the use of the polypeptide in the preparation of a medicament for treating a neurological disorder in a subject is provided, wherein the polypeptide comprises a polypeptide sequence having at least 70% sequence identity with SEQ ID NO:63 or 64.

[0189] In one embodiment, the polypeptide used according to the invention comprises a polypeptide sequence having at least 80%, 90%, 95%, or 98% sequence identity with SEQ ID NO: 63 or 64. Preferably, the polypeptide used according to the invention comprises (more preferably) the polypeptide sequence shown in SEQ ID NO: 63 or 64.

[0190] Preferably, the polypeptide containing a polypeptide sequence having at least 70% sequence identity with SEQ ID NO:63 contains a non-catalytically active L-chain, such as SEQ ID NO:64.

[0191] Chimeric and / or hybrid clostridium neurotoxins used in this invention may comprise a portion of a BoNT / A polypeptide and a portion of a BoNT / B polypeptide, examples of which include the polypeptide described herein as SEQ ID NO:44.

[0192] Suitable chimeric clostridium neurotoxins may include BoNT / FA. In fact, in a particularly preferred embodiment, the polypeptide of the present invention may comprise BoNT / FA or fragments thereof. Catalytically inactive forms of BoNT / FA are described herein as SEQ ID NO: 26 and 34. Suitable fragments of BoNT / FA are also described herein as SEQ ID NO: 28, 30, and 32.

[0193] The term "clostridium neurotoxin" can also include newly discovered members of the botulinum neurotoxin protein family expressed by non-clostridium microorganisms, such as the enterococcal toxin with the closest sequence identity to BoNT / X, the Weissella oryzae toxin called BoNT / WO (NCBI Ref. Seq: WP_027699549.1) which cleaves VAMP2 at W89-W90, the Enterococcus faecium toxin (GenBank: OT022244.1) which cleaves VAMP2 and SNAP25, and the Chryseobacterium pipero toxin (NCBI Ref. Seq: WP_034687872.1).

[0194] The polypeptides of this invention can lack the functional H of clostridium neurotoxin. C It has a structural domain and also lacks any functionally equivalent exogenous ligand targeting portion (TM).

[0195] Therefore, in a particularly preferred embodiment, the clostridial neurotoxin of the present invention is not a retargeted clostridial neurotoxin. In a retargeted clostridial neurotoxin, the clostridial neurotoxin is modified to include an exogenous ligand called a targeting moiety (TM). The TM is selected to provide binding specificity to desired target cells, and as part of the retargeting process, the natural binding moiety of the clostridial neurotoxin (e.g., H) can be removed. C domain or H CC (Structural domain). Retargeting techniques are described in, for example: EP-B-0689459; WO 1994 / 021300; EP-B-0939818; US 6,461,617; US 7,192,596; WO 1998 / 007864; EP-B-0826051; US ​​5,989,545; US 6,395,513; US 6,962,703; WO 1996 / 033273; EP-B-0996468; US 7,052,702; WO 1999 / 017806; EP-B-1107794; US 6,632,440; WO 2000 / 010598; WO 2001 / 21213; WO WO 2006 / 059093; WO 2000 / 62814; WO 2000 / 04926; WO 1993 / 15766; WO2000 / 61192; and WO 1999 / 58571; the entire contents of all these patents are incorporated herein by reference.

[0196] As described above, the (full-length) Clostridium neurotoxin consists of two polypeptide chains: a heavy chain (H chain) with a molecular weight of approximately 100 kDa and a light chain (L chain) with a molecular weight of approximately 50 kDa. The H chain contains a C-terminal targeting component (receptor-binding domain or H+). C (structural domain) and N-terminal translocation component (H) N (structural domain).

[0197] Clostridium neurotoxin may be selected from BoNT / A, BoNT / B, BoNT / C, BoNT / D, BoNT / E, BoNT / F, BoNT / G, BoNT / X, and TeNT (tetanus neurotoxin). Preferably, the clostridium neurotoxin is a botulinum neurotoxin, such as a botulinum neurotoxin selected from BoNT / A, BoNT / B, BoNT / C, BoNT / D, BoNT / E, BoNT / F, BoNT / G, and BoNT / X.

[0198] In one embodiment, the clostridial neurotoxin may be BoNT / A. The sequence of BoNT / A is shown as SEQ ID NO: 51. In another embodiment, the clostridial neurotoxin may be BoNT / B. The sequence of BoNT / B is shown as SEQ ID NO: 52. In another embodiment, the clostridial neurotoxin may be BoNT / C. The sequence of BoNT / C is shown as SEQ ID NO: 53. In another embodiment, the clostridial neurotoxin may be BoNT / D. The sequence of BoNT / D is shown as SEQ ID NO: 54. In another embodiment, the clostridial neurotoxin may be BoNT / E. The sequence of BoNT / E is shown as SEQ ID NO: 55. In another embodiment, the clostridial neurotoxin may be BoNT / F. The sequence of BoNT / F is shown as SEQ ID NO: 56. In another embodiment, the clostridial neurotoxin may be BoNT / G. The sequence of BoNT / G is shown as SEQ ID NO: 57. In another embodiment, the clostridial neurotoxin may be TeNT. The sequence of TeNT is shown as SEQ ID NO: 58. In another embodiment, the clostridial neurotoxin may be BoNT / X. The reference BoNT / X sequence is shown as SEQ ID NO:59.

[0199] In one embodiment, the polypeptide of the present invention comprises a fragment of BoNT / A or a fragment of BoNT / F. In another embodiment, the polypeptide of the present invention comprises a non-catalytically inactive L-chain of BoNT / A or BoNT / F.

[0200] In embodiments where the polypeptides described herein have tags (e.g., His-tags) and / or adapters for purification, the tags and / or adapters are optional.

[0201] This article describes suitable full-length Clostridium neurotoxins.

[0202] In one embodiment, the polypeptide of the present invention may comprise a polypeptide sequence having at least 70% sequence identity with any one of SEQ ID NO:2, 10, 12, 14, 16, 18, 26, 34, 51, 52, 53, 54, 55, 56, 57, 58, 59, 61, 62, 63, 64, or 65, provided that the clostridium neurotoxin L chain of said polypeptide is non-catalytically active. In one embodiment, the polypeptide of the present invention may comprise a polypeptide sequence having at least 80%, 90%, 95%, or 98% sequence identity with any one of SEQ ID NO:2, 10, 12, 14, 16, 18, 26, 34, 51, 52, 53, 54, 55, 56, 57, 58, 59, 61, 62, 63, 64, or 65, provided that the clostridium neurotoxin L chain of said polypeptide is non-catalytically active. Preferably, the polypeptide of the present invention may comprise a polypeptide sequence containing any one of SEQ ID NO:2, 10, 12, 14, 16, 18, 26, 34, 51, 52, 53, 54, 55, 56, 57, 58, 59, 61, 62, 63, 64 or 65, provided that the clostridium neurotoxin L chain of the polypeptide is non-catalytically active.

[0203] In one embodiment, the polypeptide of the present invention may be a polypeptide encoded by a nucleotide sequence having at least 70% sequence identity with any one of SEQ ID NO: 1, 9, 11, 13, 15, 17, 25, 33 or 60, provided that the clostridium neurotoxin L chain of the polypeptide is non-catalytically active. In another embodiment, the polypeptide of the present invention is a polypeptide encoded by a nucleotide sequence having at least 80%, 90%, 95%, or 98% sequence identity with any one of SEQ ID NO: 1, 9, 11, 13, 15, 17, 25, 33 or 60, provided that the clostridium neurotoxin L chain of the polypeptide is non-catalytically active. Preferably, the polypeptide of the present invention is a polypeptide encoded by a nucleotide sequence comprising any one of SEQ ID NO: 1, 9, 11, 13, 15, 17, 25, 33 or 60, provided that the clostridium neurotoxin L chain of the polypeptide is non-catalytically active.

[0204] In one embodiment, the polypeptide of the present invention may comprise a polypeptide sequence having at least 70% sequence identity with any one of SEQ ID NO:2, 10, 12, 14, 16, 18, 26, 34, 64, or 65, provided that the clostridium neurotoxin L-chain of the polypeptide is non-catalytically active. In one embodiment, the polypeptide of the present invention comprises a polypeptide sequence having at least 80%, 90%, 95%, or 98% sequence identity with any one of SEQ ID NO:2, 10, 12, 14, 16, 18, 26, 34, 64, or 65, provided that the clostridium neurotoxin L-chain of the polypeptide is non-catalytically active. Preferably, the polypeptide of the present invention comprises any one of SEQ ID NO:2, 10, 12, 14, 16, 18, 26, 34, 64, or 65, provided that the clostridium neurotoxin L-chain of the polypeptide is non-catalytically active.

[0205] In one embodiment, the polypeptide of the present invention is a full-length clostridium neurotoxin selected from BoNT / B, BoNT / C, BoNT / D, BoNT / E, BoNT / F, BoNT / G, BoNT / X and TeNT.

[0206] In one embodiment, the polypeptide of the present invention may comprise a polypeptide sequence having at least 70% sequence identity with any one of SEQ ID NO:52-59, 61, or 63. In one embodiment, the polypeptide of the present invention may comprise a polypeptide sequence having at least 80%, 90%, 95%, or 98% sequence identity with any one of SEQ ID NO:52-59, 61, or 63. In one embodiment, the polypeptide of the present invention may comprise a polypeptide sequence having at least 99% or 99.9% sequence identity with any one of SEQ ID NO:52-59, 61, or 63. Preferably, the polypeptide of the present invention may comprise a polypeptide sequence comprising (more preferably) any one of SEQ ID NO:52-59, 61, or 63.

[0207] In a particularly preferred embodiment, the polypeptide of the present invention is not a full-length catalytically active clostridium neurotoxin, for example, not a full-length catalytically active BoNT / A.

[0208] The polypeptides of the present invention may comprise (or consist of) fragments of clostridium neurotoxins, such as fragments of any full-length clostridium neurotoxin described herein.

[0209] In one embodiment, the polypeptide of the present invention may comprise a fragment of a polypeptide sequence having at least 70% sequence identity with any one of SEQ ID NO:2, 10, 12, 14, 16, 18, 26, 34, 51, 52, 53, 54, 55, 56, 57, 58, 59, 61, 62, 63, 64, or 65. In another embodiment, the polypeptide of the present invention may comprise a fragment of a polypeptide sequence having at least 80%, 90%, 95%, or 98% sequence identity with any one of SEQ ID NO:2, 10, 12, 14, 16, 18, 26, 34, 51, 52, 53, 54, 55, 56, 57, 58, 59, 61, 62, 63, 64, or 65. Preferably, the polypeptide of the present invention may comprise a fragment containing a polypeptide sequence of any one of SEQ ID NO: 2, 10, 12, 14, 16, 18, 26, 34, 51, 52, 53, 54, 55, 56, 57, 58, 59, 61, 62, 63, 64 or 65.

[0210] In one embodiment, the polypeptide of the present invention comprises (or consists of) a clostridial neurotoxin L-chain or a fragment thereof. The fragment of the clostridial neurotoxin L-chain may have ≤400, ≤350, ≤300, ≤250, ≤200, ≤150, ≤100, or ≤50 amino acid residues of the clostridial neurotoxin L-chain. In one embodiment, the fragment of the clostridial neurotoxin L-chain has at least 20, 30, 40, 50, 60, 70, 80, 90, 100, 120, 150, or 200 amino acid residues of the clostridial neurotoxin L-chain. For example, the fragment of the clostridial neurotoxin L-chain may have 20-400, 50-300, or 100-200 amino acid residues of the clostridial neurotoxin L-chain.

[0211] Examples of L-chain reference sequences include:

[0212] Botulinum toxin type A neurotoxin: amino acid residues 1-448

[0213] Botulinum toxin type B neurotoxin: amino acid residues 1-440

[0214] C1 botulinum neurotoxin: amino acid residues 1-441

[0215] Botulinum toxin type D neurotoxin: amino acid residues 1-445

[0216] Botulinum toxin type E neurotoxin: amino acid residues 1-422

[0217] Botulinum toxin type F neurotoxin: amino acid residues 1-439

[0218] Botulinum toxin type G neurotoxin: amino acid residues 1-441

[0219] Tetanus neurotoxin: amino acid residues 1-457

[0220] For the recently identified BoNT / X, the L-chain has been reported to correspond to its amino acid 1-439, where the L-chain boundary may vary by about 25 amino acids (e.g., 1-414 or 1-464).

[0221] The reference sequences used for identification above should be considered as guidelines, as slight variations may occur depending on the subserotype. For example, US2007 / 0166332 (incorporated hereinth by reference in its entirety) cites a slightly different Clostridium sequence:

[0222] Botulinum toxin type A neurotoxin: amino acid residue M1-K448

[0223] Botulinum toxin type B neurotoxin: amino acid residue M1-K441

[0224] C1 botulinum neurotoxin: amino acid residue M1-K449

[0225] Botulinum toxin type D neurotoxin: amino acid residue M1-R445

[0226] Botulinum toxin type E neurotoxin: Amino acid residue M1-R422

[0227] Botulinum toxin type F neurotoxin: amino acid residue M1-K439

[0228] Botulinum toxin type G neurotoxin: amino acid residue M1-K446

[0229] Tetanus neurotoxin: Amino acid residue M1-A457

[0230] This article describes a suitable Clostridium neurotoxin L chain.

[0231] The clostridial neurotoxin L-chain may comprise a polypeptide sequence having at least 70% sequence identity with any one or a fragment of SEQ ID NO: 6, 24, 32, or 40. In one embodiment, the clostridial neurotoxin L-chain comprises a polypeptide sequence having at least 80%, 90%, 95%, or 98% sequence identity with any one or a fragment of SEQ ID NO: 6, 24, 32, or 40. Preferably, the clostridial neurotoxin L-chain comprises (more preferably) a polypeptide sequence containing any one or a fragment of SEQ ID NO: 6, 24, 32, or 40.

[0232] The clostridial neurotoxin L-chain can be a chain encoded by a nucleotide sequence having at least 70% sequence identity with any one or a fragment of SEQ ID NO:5, 23, 31, or 39. In one embodiment, the clostridial neurotoxin L-chain is a chain encoded by a nucleotide sequence having at least 80%, 90%, 95%, or 98% sequence identity with any one or a fragment of SEQ ID NO:5, 23, 31, or 39. Preferably, the clostridial neurotoxin L-chain is a chain encoded by a nucleotide sequence comprising any one or a fragment of SEQ ID NO:5, 23, 31, or 39.

[0233] In one embodiment, the polypeptide of the present invention comprises (or is composed of) a fragment of the H-chain of clostridial neurotoxin. The fragment of the H-chain of clostridial neurotoxin may have ≤800, ≤700, ≤600, ≤500, ≤400, ≤350, ≤300, ≤250, ≤200, ≤150, ≤100, or ≤50 amino acid residues of the H-chain of clostridial neurotoxin. In one embodiment, the fragment of the H-chain of clostridial neurotoxin has at least 20, 30, 40, 50, 60, 70, 80, 90, 100, 120, 150, or 200 amino acid residues of the H-chain of clostridial neurotoxin. For example, the fragment of the H-chain of clostridial neurotoxin may have 20-800, 30-600, 40-400, 50-300, or 100-200 amino acid residues of the H-chain of clostridial neurotoxin.

[0234] Clostridium neurotoxin H-chain contains two structural / functional domains: a translocation domain (H... N ) and receptor-binding domain (H C ).

[0235] In one embodiment, the polypeptide of the present invention comprises a clostridium neurotoxin translocation domain or a fragment thereof (or is composed of therein). The fragment of the clostridium neurotoxin translocation domain may have ≤400, ≤350, ≤300, ≤250, ≤200, ≤150, ≤100, or ≤50 amino acid residues of the clostridium neurotoxin translocation domain. In one embodiment, the fragment of the clostridium neurotoxin translocation domain has at least 20, 30, 40, 50, 60, 70, 80, 90, 100, 120, 150, or 200 amino acid residues of the clostridium neurotoxin translocation domain. For example, the fragment of the clostridium neurotoxin translocation domain may have 20-400, 50-300, or 100-200 amino acid residues of the clostridium neurotoxin translocation domain.

[0236] The translocation domain is a fragment of the H-chain of the clostridial neurotoxin, which is approximately equivalent to the amino-terminal half of the H-chain, or corresponds to the domain of that fragment in the intact H-chain. In one embodiment, the H-chain... CThe function can be achieved by missing H C The amino acid sequence can be removed (at the DNA synthesis level, or post-synthetic level, by treatment with nucleases or proteases). Alternatively, H can be removed through chemical or biological treatment. C Functional inactivation. Therefore, in some implementations, the H-chain may not be able to bind to the binding site on the target cell where the natural clostridium neurotoxin (i.e., the whole toxin) binds.

[0237] Suitable examples of (reference) translocation domains include:

[0238] Botulinum toxin type A neurotoxin - amino acid residues (449-871)

[0239] Botulinum toxin type B neurotoxin - amino acid residues (441-858)

[0240] Botulinum toxin type C neurotoxin - amino acid residues (442-866)

[0241] Botulinum toxin type D neurotoxin - amino acid residues (446-862)

[0242] Botulinum toxin type E neurotoxin - amino acid residues (423-845)

[0243] Botulinum toxin type F neurotoxin - amino acid residues (440-864)

[0244] Botulinum toxin type G neurotoxin - amino acid residues (442-863)

[0245] Tetanus neurotoxin - amino acid residues (458-879)

[0246] The reference sequences used for identification above should be considered as guidelines, as slight variations may occur depending on the subserotype. For example, US2007 / 0166332 (incorporated herein by reference) cites a slightly different Clostridium sequence:

[0247] Botulinum toxin type A neurotoxin - amino acid residues (A449-K871)

[0248] Botulinum toxin type B neurotoxin - amino acid residues (A442-S858)

[0249] Botulinum toxin type C neurotoxin - amino acid residues (T450-N866)

[0250] Botulinum toxin type D neurotoxin - amino acid residues (D446-N862)

[0251] Botulinum toxin type E neurotoxin - amino acid residues (K423-K845)

[0252] Botulinum toxin type F neurotoxin - amino acid residues (A440-K864)

[0253] Botulinum toxin type G - amino acid residues (S447-S863)

[0254] Tetanus neurotoxin - amino acid residues (S458-V879)

[0255] In the context of this invention, various clostridium neurotoxins H containing translocation domains N The region can be used in various aspects of the present invention. In one embodiment, these active fragments can promote the release of non-cytotoxic proteases (e.g., Clostridium L-chains) from intracellular vesicles into the cytoplasm of target cells, and thus participate in the entire cellular mechanism of executing the hydrolytic cleavage of substrates by Clostridium neurotoxins. H from the heavy chain of Clostridium neurotoxins N The region is approximately 410-430 amino acids long and contains a translocation domain. Studies have shown that the H from the clostridium neurotoxin heavy chain... N The entire length of the region is not necessary for the translocation activity of the translocation domain. Therefore, various embodiments of the present invention may include clostridial neurotoxin H. N The region comprises a translocation domain having a length of, for example, at least 350 amino acids, at least 375 amino acids, at least 400 amino acids, and at least 425 amino acids. Other aspects of this embodiment may include clostridium neurotoxin H. N The region contains translocation domains having lengths of, for example, up to 350 amino acids, up to 375 amino acids, up to 400 amino acids, and up to 425 amino acids.

[0256] For further details on the genetic basis of botulinum and tetanus toxin production, see Henderson et al. (1997), The Clostridia: Molecular Biology and Pathogenesis, Academic Press.

[0257] Term H N Including naturally occurring neurotoxin H N Partially, and H modified with amino acid sequences containing amino acid residues that do not exist in nature and / or are synthetic amino acid residues. N Partial. In one implementation, the modified H N Some still exhibit the aforementioned translocation function.

[0258] In a preferred embodiment, the polypeptide of the present invention comprises a clostridium neurotoxin receptor binding domain (H... C ) or fragments thereof (or components thereof). Clostridium neurotoxin receptor-binding domain (H CFragments of this compound may contain a clostridium neurotoxin receptor binding domain (H). C The amino acid residues are ≤350, ≤300, ≤250, ≤200, ≤150, ≤100, or ≤50. In one embodiment, the clostridium neurotoxin receptor binding domain (H... C The fragment contains a clostridium neurotoxin receptor binding domain (H). C It contains at least 20, 30, 40, 50, 60, 70, 80, 90, 100, 120, 150, or 200 amino acid residues. For example, the clostridium neurotoxin receptor-binding domain (H... C Fragments of this compound may contain a clostridium neurotoxin receptor binding domain (H). C It consists of 20-350, 50-300, or 100-200 amino acid residues.

[0259] Clostridium neurotoxin receptor binding domain (H C Examples of reference sequences include:

[0260] BoNT / A-N872-L1296

[0261] BoNT / B-E859-E1291

[0262] BoNT / C1-N867-E1291

[0263] BoNT / D-S863-E1276

[0264] BoNT / E-R846-K1252

[0265] BoNT / F-K865-E1274

[0266] BoNT / G-N864-E1297

[0267] TeNT-I880-D1315

[0268] Regarding the recently identified BoNT / X, H was reported. C The domains correspond to amino acids 893-1306, with the domain boundaries possibly varying by about 25 amino acids (e.g., 868-1306 or 918-1306).

[0269] The H-chain of clostridial neurotoxin may further include a translocation-promoting domain. This domain facilitates the delivery of the L-chain into the cytoplasm of target cells and is described, for example, in WO 08 / 008803 and WO 08 / 008805, each of which is incorporated herein by reference.

[0270] For example, the translocation-promoting domain may contain clostridial neurotoxin H. CNA domain or a fragment or variant thereof. More specifically, Clostridium neurotoxin H... CN The translocation-promoting domain can have a length of at least 200 amino acids, at least 225 amino acids, at least 250 amino acids, or at least 275 amino acids. In this regard, Clostridium neurotoxin H... CN The translocation-promoting domain preferably has a length of up to 200 amino acids, up to 225 amino acids, up to 250 amino acids, or up to 275 amino acids. Specific (reference) examples include:

[0271] Botulinum toxin type A neurotoxin - amino acid residues (872-1110)

[0272] Botulinum toxin type B neurotoxin - amino acid residues (859-1097)

[0273] Botulinum toxin type C neurotoxin - amino acid residues (867-1111)

[0274] Botulinum toxin type D neurotoxin - amino acid residues (863-1098)

[0275] Botulinum toxin type E neurotoxin - amino acid residues (846-1085)

[0276] Botulinum toxin type F neurotoxin - amino acid residues (865-1105)

[0277] Botulinum toxin type G - amino acid residues (864-1105)

[0278] Tetanus neurotoxin - amino acid residues (880-1127)

[0279] Depending on the serotype / subtype, the above sequence positions may vary slightly, and appropriate (reference) Clostridium neurotoxin H CN More instances of structural domains include:

[0280] Botulinum toxin type A neurotoxin - amino acid residues (874-1110)

[0281] Botulinum toxin type B neurotoxin - amino acid residues (861-1097)

[0282] Botulinum toxin type C neurotoxin - amino acid residues (869-1111)

[0283] Botulinum toxin type D neurotoxin - amino acid residues (865-1098)

[0284] Botulinum toxin type E neurotoxin - amino acid residues (848-1085)

[0285] Botulinum toxin type F neurotoxin - amino acid residues (867-1105)

[0286] Botulinum toxin type G - amino acid residues (866-1105)

[0287] Tetanus neurotoxin - amino acid residues (882-1127)

[0288] This article describes a suitable clostridium neurotoxin H C Structural domain.

[0289] Clostridium neurotoxin H C The domain may contain a polypeptide sequence having at least 70% sequence identity with any one or a fragment of SEQ ID NO: 8, 22, 30, 38, 42, 44, 46, 48 or 50. In one embodiment, clostridium neurotoxin H C The domain comprises a polypeptide sequence having at least 80%, 90%, 95%, or 98% sequence identity with any one or a fragment of SEQ ID NO: 8, 22, 30, 38, 42, 44, 46, 48, or 50. Preferably, clostridium neurotoxin H... C The domain comprises a polypeptide sequence (more preferably composed of) any one or a fragment of SEQ ID NO:8, 22, 30, 38, 42, 44, 46, 48 or 50.

[0290] Clostridium neurotoxin H C The domain may be a domain encoded by a nucleotide sequence having at least 70% sequence identity with any one or a fragment of SEQ ID NO: 7, 21, 29, 37, 41, 43, 45, 47 or 49. In one embodiment, Clostridium neurotoxin H C The domain is encoded by a nucleotide sequence having at least 80%, 90%, 95%, or 98% sequence identity with any one or a fragment of SEQ ID NO: 7, 21, 29, 37, 41, 43, 45, 47, or 49. Preferably, clostridium neurotoxin H... C A domain is a domain encoded by a nucleotide sequence containing any one or a fragment of SEQ ID NO:7, 21, 29, 37, 41, 43, 45, 47 or 49.

[0291] In one embodiment, the clostridium neurotoxin H used in this invention C The structural domain is a variant BoNT / AHC structural domain. The variant BoNT / AHC... C The domain may contain modifications of one or more amino acid residues selected from Y1117, F1252, H1253, and L1278. For example, the variant BoNT / AH CThe structural domain may contain one or more (preferably two or more) of the following modifications: Y1117V, F1252Y, H1253K and L1278F or L1278H.

[0292] In one implementation, the variant BoNT / AH C The structural domain contains the following modifications: Y1117V and H1253K; or Y1117V, F1252Y, H1253K and L1278F; or Y1117V, F1252Y, H1253K and L1278H.

[0293] Preferred variant: BoNT / AH C The structural domain contains the following modifications: Y1117V and H1253K; or Y1117V, F1252Y, H1253K and L1278H.

[0294] Modifications can be made when compared to the unmodified BoNT / A as shown in SEQ ID NO:62, where the amino acid residue numbering is determined by comparison with SEQ ID NO:62. Since the presence of the methionine residue at position 1 of SEQ ID NO:62 is optional, those skilled in the art will consider the presence / absence of the methionine residue when determining the amino acid residue numbering. For example, if SEQ ID NO:62 contains methionine, the position numbering will be as defined above (e.g., Y1117 will be aligned with Y1117 of SEQ ID NO:62). Alternatively, if methionine is absent in SEQ ID NO:62, the amino acid residue numbering should be changed by -1 (e.g., Y1117 will be aligned with Y1116 of SEQ ID NO:52). Similar considerations apply to the presence / absence of methionine at position 1 in other polypeptide sequences described herein, and those skilled in the art will readily determine the correct amino acid residue numbering using techniques conventional in the art.

[0295] Variant BoNT / AH C The domain may contain a polypeptide sequence having at least 70% sequence identity with any one of SEQ ID NO:46, 48 or 50 or a fragment thereof, provided that it is a variant BoNT / AH. C The structural domain contains the modifications described above. In one implementation, the variant BoNT / AH C The domain comprises a polypeptide sequence having at least 80%, 90%, 95%, or 98% sequence identity with any one of SEQ ID NO:46, 48, or 50 or a fragment thereof, provided that the variant BoNT / AH is present. C The structural domain contains the modifications described above. In one implementation, the variant BoNT / AH CThe domain contains a polypeptide sequence having at least 99% or 99.9% sequence identity with any one of SEQ ID NO:46, 48 or 50 or a fragment thereof, provided that it is a variant BoNT / AH. C The structural domain contains the modifications described above. Preferably, the variant BoNT / H C The domain contains a polypeptide sequence (more preferably composed of) any one of SEQ ID NO:46, 48 or 50 or a fragment thereof.

[0296] Variant BoNT / AH C The domain may contain a polypeptide sequence having at least 70% sequence identity with any one of SEQ ID NO:46 or 50 or a fragment thereof, provided that it is a variant BoNT / AH. C The structural domain contains the modifications described above. In one implementation, the variant BoNT / AH C The domain comprises a polypeptide sequence having at least 80%, 90%, 95%, or 98% sequence identity with any one of SEQ ID NO:46 or 50 or a fragment thereof, provided that the variant BoNT / A HC domain comprises the modifications described above. In one embodiment, the variant BoNT / A HC domain... C The domain contains a polypeptide sequence having at least 99% or 99.9% sequence identity with any one of SEQ ID NO:46 or 50 or a fragment thereof, provided that it is the variant BoNT / AH. C The structural domain contains the modifications described above. Preferably, the variant BoNT / H C The domain contains a polypeptide sequence (more preferably composed of) any one or a fragment of SEQ ID NO:46 or 50.

[0297] Variant BoNT / AH C The domain may be a domain encoded by a nucleotide sequence having at least 70% sequence identity with any one of SEQ ID NO:45, 47 or 49 or a fragment thereof, provided that it is a variant BoNT / AH. C The structural domain contains the modifications described above. In one implementation, the variant BoNT / AH C The domain is encoded by a nucleotide sequence having at least 80%, 90%, 95%, or 98% sequence identity with any one of SEQ ID NO: 45, 47, or 49 or a fragment thereof, provided that it is a variant BoNT / AH. C The structural domain contains the modifications described above. In one implementation, the variant BoNT / AH CThe domain is encoded by a nucleotide sequence having at least 99% or 99.9% sequence identity with any one of SEQ ID NO:45, 47 or 49 or a fragment thereof, provided that it is a variant BoNT / AH. C The structural domain contains the modifications described above. Preferably, the variant BoNT / H C A structural domain is a structural domain encoded by any one of SEQ ID NO:45, 47 or 49 or a fragment thereof.

[0298] Variant BoNT / AH C The domain may be a domain encoded by a nucleotide sequence having at least 70% sequence identity with any of or a fragment of SEQ ID NO:45 or 49, provided that it is a variant BoNT / AH. C The structural domain contains the modifications described above. In one implementation, the variant BoNT / AH C The domain is encoded by a nucleotide sequence having at least 80%, 90%, 95%, or 98% sequence identity with any one of SEQ ID NO:45 or 49 or a fragment thereof, provided that it is a variant BoNT / AH. C The structural domain contains the modifications described above. In one implementation, the variant BoNT / AH C The domain is encoded by a nucleotide sequence having at least 99% or 99.9% sequence identity with any one of SEQ ID NO:45 or 49 or a fragment thereof, provided that it is a variant BoNT / AH. C The structural domain contains the modifications described above. Preferably, the variant BoNT / H C A structural domain is a structural domain encoded by any one of SEQ ID NO:45 or 49 or a fragment thereof.

[0299] Any of the aforementioned promoting domains can be combined with any of the aforementioned translocation domain peptides applicable to the present invention. Thus, for example, a non-clostridium promoting domain can be combined with a non-clostridium translocation domain peptide or with a clostridium translocation domain peptide. Alternatively, clostridium neurotoxin H... CN The translocation-promoting domain can combine with non-clostridium translocation domain peptides. Alternatively, clostridium neurotoxin H... CN The promoting domain can be combined with Clostridium translocation domain peptides, examples of which include:

[0300] Botulinum toxin type A neurotoxin - amino acid residues (449-1110)

[0301] Botulinum toxin type B neurotoxin - amino acid residues (442-1097)

[0302] Botulinum toxin type C neurotoxin - amino acid residues (450-1111)

[0303] Botulinum toxin type D neurotoxin - amino acid residues (446-1098)

[0304] Botulinum toxin type E neurotoxin - amino acid residues (423-1085)

[0305] Botulinum toxin type F neurotoxin - amino acid residues (440-1105)

[0306] Botulinum toxin type G - amino acid residues (447-1105)

[0307] Tetanus neurotoxin - amino acid residues (458-1127)

[0308] In some embodiments, the clostridial neurotoxin of the present invention may lack the functional H of the clostridial neurotoxin. C Structural domain. In one embodiment, the clostridial neurotoxin preferably lacks the last 50 C-terminal amino acids of the clostridial neurotoxin holotoxin. In another embodiment, the clostridial neurotoxin preferably lacks the last 100, more preferably the last 150, more preferably the last 200, particularly preferably the last 250, and most preferably the last 300 C-terminal amino acid residues of the clostridial neurotoxin holotoxin. Alternatively, H C Binding activity can be eliminated / reduced through mutagenesis—for example, for convenience, refer to BoNT / A, modifications of one or two amino acid residues in the ganglioside binding pouch (W1266 to L and Y1267 to F) result in H C The region loses its receptor-binding function. Similar mutations can be made to the non-serum type A Clostridium peptide component, for example, based on constructs of mutated Botulinum toxin B (W1262 to L and Y1263 to F) or Botulinum toxin E (W1224 to L and Y1225 to F). Other mutations at the active site achieve H C Identical elimination of receptor-binding activity, such as the Y1267S residue in botulinum toxin type A and the corresponding highly conserved residues in other clostridial neurotoxins. Details of this mutation and others are described in Rummel et al. (2004) (Molecular Microbiol. 51:631-634), which is incorporated herein by reference.

[0309] H of natural clostridium neurotoxin C Peptides contain approximately 400-440 amino acid residues and consist of two functionally distinct domains, each approximately 25 kDa, namely the N-terminal region (often called the H-terminal region). CN Peptide or structural domain) and C-terminal region (often called H) CC(peptide or domain). This fact is corroborated by the following publications, each of which is incorporated herein by reference in its entirety: Umland TC (1997) Nat. Struct. Biol. 4: 788-792; Herreros J (2000) Biochem. J. 347: 199-204; Halpern J (1993) J. Biol. Chem. 268: 15, pp. 11188-11192; Rummel A (2007) PNAS 104: 359-364; Lacey DB (1998) Nat. Struct. Biol. 5: 898-902; Knapp (1998) Am. Cryst. Assoc. Abstract Papers 25: 90; Swaminathan and Eswaramoorthy (2000) Nat. Struct. Biol. 7: 1751-1759; and Rummel A(2004)Mol.Microbiol.51(3),631-643. Furthermore, the C-terminal region (H...) has been sufficiently demonstrated. CC (Its C-terminus consists of 160-200 amino acid residues) responsible for the binding of clostridial neurotoxins to their natural cellular receptors (i.e., nerve endings at the neuromuscular junction) – a fact also confirmed by the aforementioned publications. Therefore, the lack of a functional heavy chain H is mentioned throughout this specification. C The peptide (or domain) that prevents the heavy chain from binding to cell surface receptors that naturally occur to clostridium neurotoxins means that the clostridium heavy chain simply lacks functional H. CC Peptide. In other words, H CC The peptide region may be partially or completely absent, or otherwise modified (e.g., by conventional chemical or proteolytic treatment) to reduce its natural binding ability to nerve endings at the neuromuscular junction.

[0310] Therefore, in one embodiment, the Clostridium neurotoxin H of the present invention N The peptide lacks the C-terminal peptide moiety (H) of clostridium neurotoxin. CC It is part of the body and therefore lacks the natural clostridium neurotoxin H. C Combining functions. For example, in one implementation, the C-terminus extended Clostridium H... N The peptide lacks the following C-terminal amino acid residues: 40, 60, 80, 100, 120, 140, 150, or 160 amino acid residues of the clostridium neurotoxin heavy chain. In another embodiment, the clostridium H of the present invention... NThe peptide lacks the entire C-terminal peptide moiety (H) of clostridium neurotoxin. CC And therefore lacks the H natural clostridium neurotoxin. C Combined functionality. For example, in one implementation, Clostridium H... N The peptide lacks the following C-terminal amino acid residues: 165, 170, 180, 190, or 195 amino acid residues of the clostridium neurotoxin heavy chain. As another example, the clostridium H... N Peptide deficiency is selected from the following Clostridium H CC Reference sequence:

[0311] Botulinum toxin type A neurotoxin - amino acid residues (Y1111-L1296)

[0312] Botulinum toxin type B neurotoxin - amino acid residues (Y1098-E1291)

[0313] Botulinum toxin type C neurotoxin - amino acid residues (Y1112-E1291)

[0314] Botulinum toxin type D neurotoxin - amino acid residues (Y1099-E1276)

[0315] Botulinum toxin type E neurotoxin - amino acid residues (Y1086-K1252)

[0316] Type F botulinum neurotoxin - amino acid residues (Y1106-E1274)

[0317] Botulinum toxin type G - amino acid residues (Y1106-E1297)

[0318] Tetanus neurotoxin - amino acid residues (Y1128-D1315).

[0319] The reference sequences used for identification above should be considered as guidelines, as slight variations may occur depending on the subserotype.

[0320] In a preferred embodiment, the polypeptide of the present invention comprises (or consists of) a fragment of the clostridial neurotoxin L-chain or a fragment of the clostridial neurotoxin H-chain. For example, the polypeptide may comprise a fragment of the clostridial neurotoxin L-chain or a clostridial neurotoxin translocation domain (H). N (or composed of). Preferably, the polypeptide does not further contain a clostridium neurotoxin receptor binding domain (H). C ) or Clostridium neurotoxin receptor binding domain (H CCAt least the C-terminal portion of the clostridium neurotoxin receptor binding domain (H). Therefore, in one embodiment, the polypeptide of the present invention lacks the clostridium neurotoxin receptor binding domain (H). CC The C-terminal portion of the peptide. Advantageously, such peptides lack the ability to bind to endogenous clostridium neurotoxin receptors, and therefore exhibit fewer off-target effects in subjects administered the peptide.

[0321] In one embodiment, the polypeptide of the present invention is substantially composed of a clostridial neurotoxin L-chain or a fragment thereof and / or a fragment of a clostridial neurotoxin H-chain. As used in this context, the term "substantially composed of" means that the polypeptide does not further contain one or more amino acid residues that confer additional functionality to the polypeptide, for example, when administered to a subject. In other words, a polypeptide "substantially composed of a clostridial neurotoxin L-chain or a fragment thereof and / or a fragment of a clostridial neurotoxin H-chain" may further contain one or more amino acid residues (specifically, amino acid residues targeting a clostridial neurotoxin L-chain or a fragment thereof and / or a fragment of a clostridial neurotoxin H-chain), but said one or more additional amino acid residues do not confer additional functionality to the polypeptide, for example, when administered to a subject. Additional functionality may include enzymatic activity, binding activity, and / or any physiological activity.

[0322] In one embodiment, in addition to any clostridium neurotoxin sequence, the polypeptide may also contain a non-clostridium neurotoxin sequence. The non-clostridium neurotoxin sequence preferably does not impair the ability of the polypeptide of the present invention to promote neuronal growth or neuronal repair. Preferably, the non-clostridium neurotoxin sequence is not a sequence having catalytic activity (e.g., enzyme activity). Preferably, the non-clostridium sequence is not a sequence that binds to a cell receptor. In other words, most preferably, the non-clostridium sequence is not a ligand of a cell receptor. The cell receptor may be a protein cell receptor, such as an intrinsic membrane protein. Examples of cell receptors can be found in the IUPHAR Guide to Pharmacology Database, Version 2019.4, available at https: / / www.guidetopharmacology.org / download.jsp#db_reports. The non-clostridium neurotoxin sequence may include tags that facilitate purification, such as a His tag. Preferably, any clostridium neurotoxin sequence contained in the polypeptide consists of a clostridium neurotoxin L-chain or a fragment thereof and / or a fragment of a clostridium neurotoxin H-chain. In one embodiment, the clostridium neurotoxin sequence contained in the polypeptide may consist of a clostridium neurotoxin L-chain. In one embodiment, the clostridial neurotoxin sequence contained in the polypeptide may consist of a clostridial neurotoxin translocation domain. In one embodiment, the clostridial neurotoxin sequence contained in the polypeptide may consist of a clostridial neurotoxin receptor-binding domain. In one embodiment, the clostridial neurotoxin sequence contained in the polypeptide may consist of a clostridial neurotoxin L-chain and a clostridial neurotoxin translocation domain.

[0323] This article describes suitable polypeptides containing (or composed of) the L-chain of clostridial neurotoxin and a translocation domain.

[0324] Clostridium neurotoxin comprising a clostridium neurotoxin L-chain and a translocation domain (or composed thereof) may comprise a polypeptide sequence having at least 70% sequence identity with any one or a fragment of SEQ ID NO:4, 20, 28, or 36. In one embodiment, the clostridium neurotoxin comprising a clostridium neurotoxin L-chain and a translocation domain (or composed thereof) comprises a polypeptide sequence having at least 80%, 90%, 95%, or 98% sequence identity with any one or a fragment of SEQ ID NO:4, 20, 28, or 36. Preferably, the clostridium neurotoxin comprising a clostridium neurotoxin L-chain and a translocation domain (or composed thereof) comprises a polypeptide sequence or a fragment thereof containing any one of SEQ ID NO:4, 20, 28, or 36 (more preferably composed thereof).

[0325] Clostridium neurotoxin comprising (or consisting of) a clostridium neurotoxin L-chain and a translocation domain can be a clostridium neurotoxin encoded by a nucleotide sequence having at least 70% sequence identity with any one of SEQ ID NO:3, 19, 27, or 35 or a fragment thereof. In one embodiment, the clostridium neurotoxin comprising (or consisting of) a clostridium neurotoxin L-chain and a translocation domain is a clostridium neurotoxin encoded by a nucleotide sequence having at least 80%, 90%, 95%, or 98% sequence identity with any one of SEQ ID NO:3, 19, 27, or 35 or a fragment thereof. Preferably, the clostridium neurotoxin comprising (or consisting of) a clostridium neurotoxin L-chain and a translocation domain is a clostridium neurotoxin encoded by a nucleotide sequence comprising any one of SEQ ID NO:3, 19, 27, or 35 or a fragment thereof.

[0326] The polypeptides of the present invention may not contain the complex proteins present in naturally occurring clostridium neurotoxin complexes.

[0327] The polypeptides of the present invention can be generated using recombinant nucleic acid technology. Therefore, in one embodiment, the polypeptide (as described above) is a recombinant polypeptide.

[0328] In one embodiment, a nucleic acid (e.g., DNA) comprising a nucleic acid sequence encoding a polypeptide is provided. In another embodiment, the nucleic acid sequence is prepared as part of a DNA vector comprising a promoter and a terminator.

[0329] In a preferred embodiment, the vector has a promoter selected from the following:

[0330]

[0331] In another preferred embodiment, the vector has a promoter selected from the following:

[0332]

[0333] Nucleic acid molecules can be prepared using any suitable method known in the art. Therefore, nucleic acid molecules can be prepared using chemical synthesis techniques. Alternatively, the nucleic acid molecules of this invention can be prepared using molecular biology techniques.

[0334] The DNA constructs of the present invention are preferably designed in a computer and then synthesized using conventional DNA synthesis techniques.

[0335] Depending on the final host cell (e.g., E. coli) expression system to be used, the above nucleic acid sequence information may be optionally modified according to codon preferences.

[0336] The terms "nucleotide sequence" and "nucleic acid" are used synonymously in this document. Preferably, the nucleotide sequence is a DNA sequence.

[0337] The polypeptides of the present invention (especially any of their clostridium neurotoxin portions) can exist as single strands or as double strands.

[0338] This invention provides a method for generating a single-chain polypeptide having a light chain and a heavy chain, the method comprising expressing the nucleic acid described herein in an expression host, lysing host cells to provide a host cell homogenate containing the single-chain polypeptide, and isolating the single-chain polypeptide. In one aspect, this invention provides a method for activating the polypeptide described herein, the method comprising contacting the polypeptide with a protease that hydrolyzes peptide bonds in the activation loop of the polypeptide, thereby converting the (single-chain) polypeptide into a corresponding double-chain polypeptide (e.g., wherein the light chain and heavy chain are linked together by disulfide bonds).

[0339] Therefore, the present invention provides a double-chain polypeptide that can be obtained by the method of the present invention.

[0340] Embodiments relating to various therapeutic uses of the present invention are intended to be applied equivalently to the therapeutic methods and peptides of the present invention, and vice versa.

[0341] sequence homology

[0342] Any of a variety of sequence alignment methods can be used to determine the percentage of identity, including but not limited to global methods, local methods, and hybrid methods such as segmentation methods. Schemes for determining the percentage of identity are standard procedures within the scope of those skilled in the art. Global methods align sequences from the beginning to the end of the molecule and determine the optimal alignment by accumulating the scores of individual residue pairs and by applying a vacancy penalty. Non-restrictive methods include, for example, CLUSTAL W, see Julie D. Thompson et al., CLUSTAL W: Improving the Sensitivity of Progressive Multiple Sequence Alignment Through Sequence Weighting, Position-Specific Gap Penalties and Weight Matrix Choice, 22(22) Nucleic Acids Research 4673-4680 (1994); and iterative improvements, see, for example, Osamu Gotoh, Significant Improvement in Accuracy of Multiple Protein Sequence Alignments by Iterative Refinement as Assessed by Reference to Structural Alignments, 264(4) J. MoI. Biol. 823-838 (1996). Local methods align sequences by identifying one or more conserved motifs common to all input sequences.Non-restrictive methods include, for example, match-box, see, for example, Eric Depiereux and Ernest Feytmans, Match-Box: A Fundamentally New Algorithm for the Simultaneous Alignment of Several Protein Sequences, 8(5) CABIOS 501-509 (1992); Gibbs sampling, see, for example, C.E. Lawrence et al., Detecting Subtle Sequence Signals: A Gibbs Sampling Strategy for Multiple Alignment, 262(5131) Science 208-214 (1993); Align-M, see, for example, Ivo Van Waille et al., Align-MA New Algorithm for Multiple Alignment of Highly Divergent Sequences, 20(9) Bioinformatics: 1428-1435 (2004).

[0343] Therefore, the percentage of sequence identity is determined using conventional methods. See, for example, Altschul et al., Bull. Math. Bio. 48:603-16, 1986 and Henikoff and Henikoff, Proc. Natl. Acad. Sci. USA 89:10915-19, 1992. In short, as shown below, the alignment of two amino acid sequences is optimized using a vacancy opening penalty of 10, a vacancy extension penalty of 1, and Henikoff and Henikoff's "blosum 62" scoring matrix (ibid.) to determine the alignment score (amino acids are represented by standard single-letter codes).

[0344] The "percentage of sequence identity" between two or more nucleic acid or amino acid sequences is a function of the number of common positions in the sequences. Therefore, identity % can be calculated as the number of identical nucleotides / amino acids divided by the total number of nucleotides / amino acids, then multiplied by 100. The calculation of % sequence identity can also take into account the number of vacancies that need to be introduced to optimize the alignment of two or more sequences, as well as the length of each vacancy. Specific mathematical algorithms familiar to those skilled in the art (e.g., BLAST) can be used for sequence comparison and to determine the percentage of identity between two or more sequences.

[0345] Alignment score for determining sequence identity

[0346]

[0347] Then, the percentage identity is calculated as follows:

[0348]

[0349] Essentially homologous peptides are characterized by having one or more amino acid substitutions, deletions, or additions. These changes are preferably insignificant, i.e., conserved amino acid substitutions (see below) and other substitutions that do not significantly affect peptide folding or activity; small deletions, typically 1 to about 30 amino acids; and small amino or carboxyl terminal extensions, such as N-terminal methionine residues, or small linker peptides or affinity tags of up to about 20-25 residues.

[0350] Conservative amino acid substitution

[0351] Alkaline: Arginine

[0352] Lysine

[0353] Histidine

[0354] Acidity: Glutamic acid

[0355] Aspartic acid

[0356] Polarity: glutamine

[0357] Asparagine

[0358] Hydrophobicity: Leucine

[0359] Isoleucine

[0360] Valine

[0361] Aromatic compounds: Phenylalanine

[0362] Tryptophan

[0363] Tyrosine

[0364] Small: Glycine

[0365] alanine

[0366] Serine

[0367] threonine

[0368] Methionine

[0369] In addition to the 20 standard amino acids, non-standard amino acids (e.g., 4-hydroxyproline, 6-N-methyllysine, 2-aminoisobutyric acid, isovaleine, and α-methylserine) may replace amino acid residues in the polypeptides of the present invention. A limited number of non-conserved amino acids, amino acids not encoded by the genetic code, and non-natural amino acids may replace amino acid residues in the polypeptides. The polypeptides of the present invention may also contain non-naturally occurring amino acid residues.

[0370] Non-naturally occurring amino acids include, but are not limited to, trans-3-methylproline, 2,4-methylproline, cis-4-hydroxyproline, trans-4-hydroxyproline, N-methylglycine, allothreonine, methylthreonine, hydroxyethylcysteine, hydroxyethylhomocysteine, nitroglutamine, homoglutamine, piperidine acid, tert-leucine, valine, 2-azaphenylalanine, 3-azaphenylalanine, 4-azaphenylalanine, and 4-fluorophenylalanine. Several methods for incorporating non-naturally occurring amino acid residues into proteins are known in the art. For example, an in vitro system can be used in which nonsense mutations are suppressed using chemically aminoacylated repressor tRNAs. Methods for synthesizing amino acids and aminoacylated tRNAs are known in the art. Transcription and translation of plasmids containing nonsense mutations are performed in a cell-free system containing *E. coli* S30 extract and commercially available enzymes and other reagents. The protein is purified by chromatography. See, for example, Robertson et al., J. Am. Chem. Soc. 113:2722, 1991; Ellman et al., Methods Enzymol. 202:301, 1991; Chung et al., Science 259:806-9, 1993; and Chung et al., Proc. Natl. Acad. Sci. USA 90:10145-9, 1993). In the second method, translation is performed in Xenopus oocytes by microinjection of mutated mRNA and chemically aminoacylated repressor tRNA (Turcatti et al., J. Biol. Chem. 271:19991-8, 1996). In the third method, *E. coli* cells are cultured in the absence of the natural amino acid to be substituted (e.g., phenylalanine) and in the presence of the desired non-natural amino acid (e.g., 2-azaphenylalanine, 3-azaphenylalanine, 4-azaphenylalanine, or 4-fluorophenylalanine). The non-natural amino acid is incorporated into the polypeptide, replacing its natural counterpart. See Koide et al., Biochem. 33:7470-6, 1994. Naturally occurring amino acid residues can be converted into non-natural substances through in vitro chemical modification. Chemical modification can be used in combination with site-directed mutagenesis to further expand the range of substitutions (Wynn and Richards, Protein Sci. 2:395-403, 1993).

[0371] A limited number of non-conserved amino acids, amino acids not encoded by the genetic code, non-naturally occurring amino acids, and non-natural amino acids can replace the amino acid residues of the polypeptides of the present invention.

[0372] Essential amino acids in the polypeptides of the present invention can be identified according to procedures known in the art, such as site-directed mutagenesis or alanine scanning mutagenesis (Cunningham and Wells, Science 244:1081-5, 1989). Sites of biological interaction can also be determined by physical analysis of the structure, such as by techniques like nuclear magnetic resonance, crystallography, electron diffraction, or photoaffinity labeling, combined with assumed contact site amino acid mutations. See, for example, de Vos et al., Science 255:306-12, 1992; Smith et al., J. Mol. Biol. 224:899-904, 1992; Wlodaver et al., FEBS Lett. 309:59-64, 1992. The identification of essential amino acids can also be inferred from homology analysis of related components (e.g., translocation or protease components) of the polypeptides of the present invention.

[0373] Multiple amino acid substitutions can be performed and tested using known methods of mutagenesis and screening, such as those disclosed in Reidhaar-Olson and Sauer (Science 241:53-7, 1988) or Bowie and Sauer (Proc. Natl. Acad. Sci. USA 86:2152-6, 1989). In short, these authors disclosed methods for simultaneously randomizing two or more positions in a polypeptide, selecting functional polypeptides, and then sequencing the mutagenized polypeptide to determine the range of permissible substitutions at each position. Other methods that can be used include phage display (e.g., Lowman et al., Biochem. 30:10832-7, 1991; Ladner et al., U.S. Patent No. 5,223,409; Huse, WIPO Publication WO92 / 06204) and region-directed mutagenesis (Derbyshire et al., Gene 46:145, 1986; Ner et al., DNA 7:127, 1988).

[0374] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains. Singleton et al., DICTIONARY OF MICROBIOLOGY AND MOLECULAR BIOLOGY, 20th edition, John Wiley and Sons, New York (1994), and Hale & Marham, THE HARPER COLLINS DICTIONARY OF BIOLOGY, Harper Perennial, NY (1991), provide a general dictionary for skilled workers of the subject of the art of the many terms used in this disclosure.

[0375] This disclosure is not limited to the exemplary methods and materials disclosed herein, and any methods and materials similar to or equivalent to those described herein may be used in the practice or testing of embodiments of this disclosure. Numerical ranges include the numbers defining the range. Unless otherwise stated, any nucleic acid sequence is written from left to right in a 5' to 3' orientation; amino acid sequences are written from left to right, respectively, from amino to carboxyl.

[0376] The headings provided herein are not intended to limit any aspect or implementation of this disclosure.

[0377] In this document, amino acid names, three-letter abbreviations, or single-letter abbreviations are used to refer to amino acids. As used herein, the term "protein" includes proteins, polypeptides, and peptides. As used herein, the term "amino acid sequence" is synonymous with the terms "polypeptide" and / or "protein." In some cases, the term "amino acid sequence" is synonymous with the term "peptide." In some cases, the term "amino acid sequence" is synonymous with the term "enzyme." The terms "protein" and "polypeptide" are used interchangeably herein. Conventional single-letter and three-letter codons for amino acid residues may be used in this disclosure and claims. Three-letter codons for amino acids are defined according to the Joint Committee on Biochemical Nomenclature of IUPACIUB (JCBN). It should also be understood that, due to the degeneracy of the genetic codon, a polypeptide can be encoded by more than one nucleotide sequence.

[0378] Other definitions of terms may appear throughout the specification. Before describing the exemplary embodiments in more detail, it should be understood that this disclosure is not limited to the specific embodiments described and therefore can vary. It should also be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting, as the scope of this disclosure is defined only by the appended claims.

[0379] In the case of a range of values ​​provided, it should be understood that, unless the context explicitly indicates otherwise, each midpoint between the upper and lower limits of the range, up to one-tenth of the lower limit unit, is specifically included in this disclosure. Each smaller range between any specified value or midpoint within the specified range and any other specified value or midpoint within that specified range is included in this disclosure. The upper and lower limits of these smaller ranges may be independently included or excluded from the range, and each range that includes one or both of the upper and lower limits of the smaller range, excludes neither, or includes both is also included in this disclosure, based on any explicitly excluded limit within the range. Where the range includes one or both limits, ranges that exclude any one or both of those included limits are also included in this disclosure.

[0380] It is important to note that, as used herein and in the appended claims, the singular forms “a,” “an,” and “the” include plural indicators unless the context clearly indicates otherwise. Thus, for example, reference to “clostridium neurotoxin” includes a variety of such candidate agents, and reference to “clostridium neurotoxin” includes reference to one or more clostridium neurotoxins and their equivalents known to those skilled in the art, etc.

[0381] The publications discussed herein are provided solely because their contents were disclosed prior to the filing date of this application. Nothing herein should be construed as an admission that such publications constitute prior art as claimed in the appended claims. Attached Figure Description

[0382] Schemes of the present invention will now be described by way of example only with reference to the following figures and embodiments.

[0383] Figure 1 This study demonstrates the neurotrophic effects of different recombinant expressions of non-catalytically active BoNT serotypes compared to the positive control brain-derived neurotrophic factor (BDNF) in the motor neuron-like cell line NSC34. *p<0.05 relative to the untreated control, one-way ANOVA followed by Dunnett's multiple comparison test. Data are the mean ± standard error of three independent experiments, each performed in six replicate wells.

[0384] Figure 2 This study demonstrates the neurotrophic effects of botulinum neurotoxin serotype A fragment in the motor neuron-like cell line NSC34 and the effects of recombinantly expressed, non-catalytically active BoNT / A. BDNF was used as a positive control. *p<0.05 relative to untreated controls, one-way ANOVA followed by Dunnett's multiple comparison test. Data are the mean ± standard error of three independent experiments, each performed in six replicate wells.

[0385] Figure 3 The neurotrophic effects of negative control and recombinant expression of catalytically inactive BoNT / A (BoNT / A(0)) in the motor neuron-like cell line NSC34 were shown. BDNF was used as a positive control. *p<0.05 relative to the untreated control, one-way ANOVA followed by Dunnett's multiple comparison test. Data are mean ± standard error of three independent experiments, each performed in six replicate wells.

[0386] Figure 4 The results of the horizontal ladder test are shown in mice that were administered the load control (PBS) or 100 pg, 100 ng or 50 ug of rBoNT / A(0).

[0387] Figure 5 The following are shown: (A) Immunohistochemistry using an antibody bound to neurofilament 200 (NF200) 4 weeks after administration of the carrier (PBS) (left panel) or 100 ng rBoNT / A(0) (right panel); and (B) Immunohistochemistry using an antibody bound to MAP1B 4 weeks after administration of the carrier (PBS) (left panel) or 100 ng rBoNT / A(0) (right panel). Lesion sites are indicated by * (and for...). Figure 5 B, indicated by a white arrow).

[0388] Figure 6 The images show (A) a catalytically inactive BoNT / A(0) fragment and (B) a BoNT / A light chain addition translocation domain fragment (LH). N (A), (C)BoNT / A light chain (LC / A, i.e., L / A), and (D)BoNT / A receptor-binding domain (H C / A) Effect on the number of neurites per cell. BoNT or BoNT fragments were compared with BSA (negative control) and BDNF (positive control) at concentrations of 0.1 nM, 1 nM, and 10 nM. *p < 0.05 relative to the BSA control. One-way ANOVA was performed, followed by Dunnett's post hoc test. Data are presented as mean ± SE.

[0389] Figure 7 The images show (A) a catalytically inactive BoNT / FA (0) fragment and (B) a BoNT / FA light chain addition translocation domain fragment (LH). N (C)BoNT / FA light chain (LC / FA, i.e., L / FA) and (D)BoNT / FA receptor-binding domain (H) CThe effect of BoNT or BoNT fragments on the number of neurites per cell. BoNT or BoNT fragments were compared with BSA (negative control) and BDNF (positive control) at concentrations of 0.1 nM, 1 nM, and 10 nM. *p < 0.05 relative to the BSA control. One-way ANOVA followed by Dunnett's post-hoc test. Data are presented as mean ± SE.

[0390] Figure 8 This shows a fragment of (A)BoNT / F light chain plus translocation domain (LH). N (B) BoNT / F light chain (LC / F, i.e., L / F) and (C) BoNT / F receptor-binding domain (H C The effect of BoNT or BoNT fragments on the number of neurites per cell. BoNT or BoNT fragments were compared with BSA (negative control) and BDNF (positive control) at concentrations of 0.1 nM, 1 nM, and 10 nM. *p < 0.05 relative to the BSA control. One-way ANOVA followed by Dunnett's post-hoc test. Data are presented as mean ± SE.

[0391] Figure 9 The rH of the cation was shown C The effect of mrHC / A (i.e., mrHC / A) on the number of neurites per cell. The BoNT fragment of the cation was compared with BSA (negative control) and BDNF (positive control) at concentrations of 0.1 nM, 1 nM, and 10 nM. *p < 0.05 relative to the BSA control. One-way ANOVA was performed, followed by Dunnett's post-hoc test. Data are presented as mean ± SE.

[0392] Figure 10 It shows (A)toxHC / A YH (i.e., rH) C (A) variant Y1117V H1253K) and (B) toxHC / A YFHL(L to H) (i.e. rH) C The effect of variant BoNT fragment Y1117V F1252Y H1253K L1278H on the number of neurites per cell. The BoNT variant fragment was compared with BSA (negative control) and BDNF (positive control) at concentrations of 0.1 nM, 1 nM, and 10 nM. *p < 0.05 relative to the BSA control. One-way ANOVA followed by Dunnett's post-hoc test was performed. Data are presented as mean ± SE. Figure 11 The experimental protocol used for screening on the neuronal cell line (NSC34) is shown.

[0393] Specific implementation of the case

[0394] 1. A polypeptide for promoting neuronal growth or neuronal repair to treat neurological disorders in a subject, wherein the polypeptide comprises:

[0395] Clostridium neurotoxin light chain (L-chain) or fragments thereof; and / or

[0396] Fragments of the clostridium neurotoxin heavy chain (H-chain).

[0397] 2. A method for promoting neuronal growth or neuronal repair to treat a neurological disorder in a subject, the method comprising administering a polypeptide to the subject, wherein the polypeptide comprises:

[0398] Clostridium neurotoxin L-chain or fragments thereof; and / or

[0399] Fragments of the H-chain of Clostridium neurotoxin.

[0400] 3. Use of the polypeptide in the preparation of a medicament for the treatment of neurological disorders in a subject, wherein the polypeptide comprises:

[0401] Clostridium neurotoxin L-chain or fragments thereof; and / or

[0402] Fragments of the H-chain of Clostridium neurotoxin.

[0403] 4. The polypeptide used according to embodiment 1, the method according to embodiment 2, or the use according to embodiment 3, wherein the L-chain is non-catalytically active.

[0404] 5. The polypeptide, method, or use according to any of the preceding embodiments, wherein the polypeptide is substantially composed of a clostridium neurotoxin light chain (L-chain) or a fragment thereof; and / or a fragment of a clostridium neurotoxin heavy chain (H-chain).

[0405] 6. The polypeptide, method, or use according to any of the preceding embodiments, wherein the polypeptide comprises a clostridium neurotoxin light chain (L-chain) or a fragment thereof; and / or a fragment of a clostridium neurotoxin heavy chain (H-chain).

[0406] 7. The polypeptide, method, or use according to any of the foregoing embodiments, wherein the fragment of the clostridium neurotoxin H-chain comprises: a translocation domain (H... N ) or fragments thereof; or Clostridium neurotoxin receptor binding domain (H C (or fragments thereof)

[0407] 8. The polypeptide, method, or use according to any of the foregoing embodiments, wherein the fragment of the clostridial neurotoxin H-chain contains H. N A structural domain or a fragment thereof.

[0408] 9. The polypeptide, method, or use according to any of the foregoing embodiments, wherein the fragment of the H-chain of the clostridial neurotoxin is composed of H... N Composed of a structural domain or its fragments.

[0409] 10. The polypeptide, method, or use according to any of the foregoing embodiments, wherein the fragment of the clostridium neurotoxin H-chain contains H C A structural domain or a fragment thereof.

[0410] 11. The polypeptide, method, or use according to any of the foregoing embodiments, wherein the fragment of the H-chain of the clostridium neurotoxin is composed of H... C Composed of a structural domain or its fragments.

[0411] 12. The polypeptide, method, or use according to any of the foregoing embodiments, wherein the polypeptide lacks the clostridium neurotoxin receptor-binding domain (H). CC The C-terminal portion of ).

[0412] 13. The polypeptide, method, or use according to any of the foregoing embodiments, wherein the polypeptide does not contain clostridium neurotoxin H. N Domain and H C Both structural domains.

[0413] 14. The polypeptide, method, or use according to any of the foregoing embodiments, wherein the polypeptide does not further comprise a non-clostridium catalytic domain.

[0414] 15. The polypeptide, method, or use according to any of the preceding embodiments, wherein the polypeptide comprises: a clostridium neurotoxin L-chain or a fragment thereof, and H... N A structural domain or a fragment thereof.

[0415] 16. The polypeptide, method, or use according to any of the preceding embodiments, wherein the polypeptide comprises: a clostridium neurotoxin L-chain or a fragment thereof, and H... N A structural domain or a fragment thereof.

[0416] 17. The polypeptide, method, or use according to any of the preceding embodiments, wherein the polypeptide comprises: a clostridium neurotoxin L-chain and H... N Structural domain.

[0417] 18. The polypeptide, method, or use according to any of the preceding embodiments, wherein the polypeptide:

[0418] a. Encoded by a nucleotide sequence having at least 70% sequence identity with any one of SEQ ID NO: 3, 5, 7, 19, 21, 23, 27, 29, 31, 35, 37, 39, 41, 43, 45, 47 or 49; or

[0419] b. Contains a polypeptide sequence (preferably composed of) having at least 70% sequence identity with any one of SEQ ID NO: 4, 6, 8, 20, 22, 24, 28, 30, 32, 36, 38, 40, 42, 44, 46, 48 or 50.

[0420] 19. The polypeptide, method, or use according to any of the preceding embodiments, wherein the polypeptide:

[0421] a. Encoded by a nucleotide sequence having at least 80% sequence identity with any one of SEQ ID NO: 3, 5, 7, 19, 21, 23, 27, 29, 31, 35, 37, 39, 41, 43, 45, 47 or 49; or

[0422] b. Contains a polypeptide sequence (preferably composed of) having at least 80% sequence identity with any one of SEQ ID NO: 4, 6, 8, 20, 22, 24, 28, 30, 32, 36, 38, 40, 42, 44, 46, 48 or 50.

[0423] 20. The polypeptide, method, or use according to any of the preceding embodiments, wherein the polypeptide:

[0424] a. Encoded by a nucleotide sequence having at least 90% sequence identity with any one of SEQ ID NO: 3, 5, 7, 19, 21, 23, 27, 29, 31, 35, 37, 39, 41, 43, 45, 47 or 49; or

[0425] b. Contains a polypeptide sequence (preferably composed of) having at least 90% sequence identity with any one of SEQ ID NO: 4, 6, 8, 20, 22, 24, 28, 30, 32, 36, 38, 40, 42, 44, 46, 48 or 50.

[0426] 21. The polypeptide, method, or use according to any of the foregoing embodiments, wherein the polypeptide:

[0427] a. Encoded by a nucleotide sequence having at least 95% sequence identity with any one of SEQ ID NO: 3, 5, 7, 19, 21, 23, 27, 29, 31, 35, 37, 39, 41, 43, 45, 47 or 49; or

[0428] b. Contains a polypeptide sequence (preferably composed of) having at least 95% sequence identity with any one of SEQ ID NO: 4, 6, 8, 20, 22, 24, 28, 30, 32, 36, 38, 40, 42, 44, 46, 48 or 50.

[0429] 22. The polypeptide, method, or use according to any of the foregoing embodiments, wherein the polypeptide:

[0430] a. Encoded by a nucleotide sequence having at least 99% sequence identity with any one of SEQ ID NO: 3, 5, 7, 19, 21, 23, 27, 29, 31, 35, 37, 39, 41, 43, 45, 47 or 49; or

[0431] b. Contains a polypeptide sequence (preferably composed of) having at least 99% sequence identity with any one of SEQ ID NO:4, 6, 8, 20, 22, 24, 28, 30, 32, 36, 38, 40, 42, 44, 46, 48 or 50.

[0432] 23. The polypeptide, method, or use according to any of the foregoing embodiments, wherein the polypeptide:

[0433] a. Encoded by a nucleotide sequence having at least 99.9% sequence identity with any one of SEQ ID NO: 3, 5, 7, 19, 21, 23, 27, 29, 31, 35, 37, 39, 41, 43, 45, 47 or 49; or

[0434] b. Containing (preferably composed of a polypeptide sequence having at least 99.9% sequence identity with any one of SEQ ID NO:4, 6, 8, 20, 22, 24, 28, 30, 32, 36, 38, 40, 42, 44, 46, 48 or 50).

[0435] 24. A polypeptide for promoting neuronal growth or neuronal repair to treat neurological disorders in a subject, wherein the polypeptide comprises a non-catalytically active clostridium neurotoxin L-chain.

[0436] 25. A method for promoting neuronal growth or neuronal repair to treat a subject's neurological disorder, the method comprising administering a polypeptide to the subject, wherein the polypeptide comprises a non-catalytically active clostridium neurotoxin L-chain.

[0437] 26. Use of a polypeptide containing a non-catalytically active clostridium neurotoxin L-chain in the preparation of a medicament for promoting neuronal growth or neuronal repair to treat neurological disorders in a subject.

[0438] 27. A polypeptide for promoting neuronal growth or neuronal repair to treat a subject’s neurological disorder, wherein the polypeptide comprises a polypeptide sequence having at least 70% sequence identity with SEQ ID NO:42 and / or wherein the polypeptide comprises a polypeptide sequence encoded by a nucleotide sequence having at least 70% sequence identity with SEQ ID NO:41.

[0439] 28. A method for promoting neuronal growth or neuronal repair to treat a subject's neurological disorder, the method comprising administering a polypeptide to the subject, wherein the polypeptide comprises a polypeptide sequence having at least 70% sequence identity with SEQ ID NO:42 and / or wherein the polypeptide comprises a polypeptide sequence encoded by a nucleotide sequence having at least 70% sequence identity with SEQ ID NO:41.

[0440] 29. Use of a polypeptide in the preparation of a medicament for promoting neuronal growth or neuronal repair to treat a subject’s neurological disorder, wherein the polypeptide comprises a polypeptide sequence having at least 70% sequence identity with SEQ ID NO:42 and / or wherein the polypeptide comprises a polypeptide sequence encoded by a nucleotide sequence having at least 70% sequence identity with SEQ ID NO:41.

[0441] 30. The polypeptide, method, or use according to any one of embodiments 27-29, wherein the polypeptide comprises a polypeptide sequence having at least 80% sequence identity with SEQ ID NO:42 (preferably composed of therefrom) and / or wherein the polypeptide is encoded by a nucleotide sequence having at least 80% sequence identity with SEQ ID NO:41.

[0442] 31. The polypeptide, method, or use according to any one of embodiments 27-30, wherein the polypeptide comprises a polypeptide sequence having at least 90% sequence identity with SEQ ID NO:42 (preferably composed of therefrom) and / or wherein the polypeptide is encoded by a nucleotide sequence having at least 90% sequence identity with SEQ ID NO:41.

[0443] 32. The polypeptide, method, or use according to any one of embodiments 27-31, wherein the polypeptide comprises a polypeptide sequence having at least 95% sequence identity with SEQ ID NO:42 (preferably composed of therefrom) and / or wherein the polypeptide is encoded by a nucleotide sequence having at least 95% sequence identity with SEQ ID NO:41.

[0444] 33. The polypeptide, method, or use according to any one of embodiments 27-32, wherein the polypeptide comprises a polypeptide sequence having at least 99% sequence identity with SEQ ID NO:42 (preferably composed of therefrom) and / or wherein the polypeptide is encoded by a nucleotide sequence having at least 99% sequence identity with SEQ ID NO:41.

[0445] 34. The polypeptide, method, or use according to any one of embodiments 27-33, wherein the polypeptide comprises a polypeptide sequence having at least 99.9% sequence identity with SEQ ID NO:42 (preferably composed of therefrom) and / or wherein the polypeptide is encoded by a nucleotide sequence having at least 99.9% sequence identity with SEQ ID NO:41.

[0446] 35. The polypeptide, method, or use according to any one of embodiments 27-34, wherein the polypeptide comprises a polypeptide sequence (preferably composed of) having at least 70% sequence identity with SEQ ID NO: 61 or 65 and / or wherein the polypeptide is encoded by a nucleotide sequence having at least 70% sequence identity with SEQ ID NO: 60.

[0447] 36. The polypeptide, method, or use according to any one of embodiments 27-35, wherein the polypeptide comprises a polypeptide sequence (preferably composed of) having at least 80% sequence identity with SEQ ID NO: 61 or 65 and / or wherein the polypeptide is encoded by a nucleotide sequence having at least 80% sequence identity with SEQ ID NO: 60.

[0448] 37. The polypeptide, method, or use according to any one of embodiments 27-36, wherein the polypeptide comprises a polypeptide sequence (preferably composed of) having at least 90% sequence identity with SEQ ID NO: 61 or 65 and / or wherein the polypeptide is encoded by a nucleotide sequence having at least 90% sequence identity with SEQ ID NO: 60.

[0449] 38. The polypeptide, method, or use according to any one of embodiments 27-37, wherein the polypeptide comprises a polypeptide sequence having at least 95% sequence identity with SEQ ID NO: 61 or 65 (preferably composed of therefrom) and / or wherein the polypeptide is encoded by a nucleotide sequence having at least 95% sequence identity with SEQ ID NO: 60.

[0450] 39. The polypeptide, method, or use according to any one of embodiments 27-38, wherein the polypeptide comprises a polypeptide sequence having at least 99% sequence identity with SEQ ID NO: 61 or 65 (preferably composed of therefrom) and / or wherein the polypeptide is encoded by a nucleotide sequence having at least 99% sequence identity with SEQ ID NO: 60.

[0451] 40. The polypeptide, method, or use according to any one of embodiments 27-39, wherein the polypeptide comprises a polypeptide sequence having at least 99.9% sequence identity with SEQ ID NO: 61 or 65 (preferably composed of therefrom) and / or wherein the polypeptide is encoded by a nucleotide sequence having at least 99.9% sequence identity with SEQ ID NO: 60.

[0452] 41. A polypeptide for promoting neuronal growth or neuronal repair to treat a subject’s neurological disorder, wherein the polypeptide comprises a polypeptide sequence having at least 70% sequence identity with SEQ ID NO:63 or 64.

[0453] 42. A method for promoting neuronal growth or neuronal repair to treat a subject’s neurological disorder, the method comprising administering a polypeptide to the subject, wherein the polypeptide comprises a polypeptide sequence having at least 70% sequence identity with SEQ ID NO:63 or 64.

[0454] 43. Use of a polypeptide in the preparation of a medicament for promoting neuronal growth or neuronal repair to treat a subject’s neurological disorder, wherein the polypeptide comprises a polypeptide sequence having at least 70% sequence identity with SEQ ID NO: 63 or 64.

[0455] 44. The polypeptide, method, or use according to any one of embodiments 41-43, wherein the polypeptide comprises (preferably consists of) a polypeptide sequence having at least 80% sequence identity with SEQ ID NO: 63 or 64.

[0456] 45. The polypeptide, method, or use according to any one of embodiments 41-44, wherein the polypeptide comprises (preferably consists of) a polypeptide sequence having at least 90% sequence identity with SEQ ID NO: 63 or 64.

[0457] 46. ​​The polypeptide, method, or use according to any one of embodiments 41-45, wherein the polypeptide comprises (preferably consists of) a polypeptide sequence having at least 95% sequence identity with SEQ ID NO: 63 or 64.

[0458] 47. The polypeptide, method, or use according to any one of embodiments 41-46, wherein the polypeptide comprises (preferably consists of) a polypeptide sequence having at least 99% sequence identity with SEQ ID NO: 63 or 64.

[0459] 48. The polypeptide, method, or use according to any one of embodiments 41-47, wherein the polypeptide comprises (preferably consists of) a polypeptide sequence having at least 99.9% sequence identity with SEQ ID NO: 63 or 64.

[0460] 49. The polypeptide, method, or use according to any of the preceding embodiments, wherein the polypeptide does not contain the H-chain of the natural clostridium neurotoxin.

[0461] 50. The polypeptide, method, or use according to any of the preceding embodiments, wherein the polypeptide is neurotrophic.

[0462] 51. The polypeptide, method, or use according to any of the preceding embodiments, wherein the polypeptide promotes neuronal growth and / or neuronal repair.

[0463] 52. The polypeptide, method, or use according to any of the preceding embodiments, wherein the neurological disorder is a disorder that can be treated by promoting neuronal growth and / or repair.

[0464] 53. The polypeptide, method, or use according to any of the preceding embodiments, wherein the neurological disorder is neuronal damage, neurodegenerative disorder, sensory disorder, or autonomic disorder.

[0465] 54. The polypeptide, method, or use according to any of the preceding embodiments, wherein the neurological disorder is selected from neuronal damage including: neurological trauma (e.g., caused by scarring and / or fracture), neuropathy (e.g., peripheral neuropathy), spinal cord injury (e.g., including paralysis), nerve transection, brain injury (e.g., traumatic brain injury), non-traumatic injury (e.g., stroke or spinal cord infarction), and brachial plexus injury, such as Erb's paralysis or Klumpke's paralysis.

[0466] 55. The polypeptide, method, or use according to any of the preceding embodiments, wherein the neurological disorder is selected from the following neurodegenerative disorders: Alzheimer's disease, Parkinson's disease, Parkinson's disease-related conditions, motor neuron disease, peripheral neuropathy, motor neuropathy, prions, Huntington's disease, spinocerebellar ataxia, spinal muscular atrophy, unilateral muscular atrophy, Friedreich's ataxia, Hallervorden-Spatz disease, or frontotemporal degeneration.

[0467] 56. The polypeptide, method, or use according to any of the preceding embodiments, wherein the polypeptide promotes the growth or repair of motor neurons.

[0468] 57. The polypeptide, method, or use according to any of the preceding embodiments, wherein the polypeptide is a modified clostridium neurotoxin, such as a chimeric clostridium neurotoxin or a hybrid clostridium neurotoxin.

[0469] 58. The polypeptide, method, or use according to any one of embodiments 24-34 or 49-57, wherein the polypeptide is non-catalytically active, and:

[0470] a. Encoded by a nucleotide sequence having at least 70% sequence identity with any one of SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49 or 60; or

[0471] b. A polypeptide sequence (preferably composed of) having at least 70% sequence identity with any one of SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 61, 62, 63, 64 or 65.

[0472] 59. The polypeptide, method, or use according to any one of embodiments 24-34 or 49-58, wherein the polypeptide is non-catalytically active, and:

[0473] a. Encoded by a nucleotide sequence having at least 80% sequence identity with any one of SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49 or 60; or

[0474] b. A polypeptide sequence (preferably composed of) having at least 80% sequence identity with any one of SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 61, 62, 63, 64 or 65.

[0475] 60. The polypeptide, method, or use according to any one of embodiments 24-34 or 49-59, wherein the polypeptide is non-catalytically active, and:

[0476] a. Encoded by a nucleotide sequence having at least 90% sequence identity with any one of SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49 or 60; or

[0477] b. A polypeptide sequence (preferably composed of) having at least 90% sequence identity with any one of SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 61, 62, 63, 64 or 65.

[0478] 61. The polypeptide, method, or use according to any one of embodiments 24-34 or 49-60, wherein the polypeptide is non-catalytically active, and:

[0479] a. Encoded by a nucleotide sequence having at least 95% sequence identity with any one of SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49 or 60; or

[0480] b. Containing (preferably consisting of) a polypeptide sequence having at least 95% sequence identity with any one of SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 61, 62, 63, 64 or 65.

[0481] 62. The polypeptide, method, or use according to any one of embodiments 24-34 or 49-61, wherein the polypeptide is non-catalytically active, and:

[0482] a. Encoded by a nucleotide sequence having at least 99% sequence identity with any one of SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49 or 60; or

[0483] b. A polypeptide sequence comprising (preferably consisting of) at least 99% sequence identity with any one of SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 61, 62, 63, 64, or 65.

[0484] 63. The polypeptide, method, or use according to any one of embodiments 24-34 or 49-62, wherein the polypeptide is non-catalytically active, and:

[0485] a. Encoded by a nucleotide sequence having at least 99.9% sequence identity with any one of SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49 or 60; or

[0486] b. A polypeptide sequence (preferably composed of) having at least 99.9% sequence identity with any one of SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 61, 62, 63, 64 or 65.

[0487] 64. The polypeptide, method, or use according to any one of embodiments 24-26 or 49-63, wherein the polypeptide:

[0488] a. Encoded by a nucleotide sequence having at least 70% sequence identity with any one of SEQ ID NO: 1, 9, 11, 13, 15, 17, 25 or 33; or

[0489] b. Contains a polypeptide sequence (preferably composed of) having at least 70% sequence identity with any one of SEQ ID NO:2, 10, 12, 14, 16, 18, 26, 34, 64 or 65.

[0490] 65. The polypeptide, method, or use according to any one of embodiments 24-26 or 49-64, wherein the polypeptide:

[0491] a. Encoded by a nucleotide sequence having at least 80% sequence identity with any one of SEQ ID NO: 1, 9, 11, 13, 15, 17, 25 or 33; or

[0492] b. Contains a polypeptide sequence (preferably composed of) having at least 80% sequence identity with any one of SEQ ID NO:2, 10, 12, 14, 16, 18, 26, 34, 64 or 65.

[0493] 66. The polypeptide, method, or use according to any one of embodiments 24-26 or 49-65, wherein the polypeptide:

[0494] a. Encoded by a nucleotide sequence having at least 90% sequence identity with any one of SEQ ID NO: 1, 9, 11, 13, 15, 17, 25 or 33; or

[0495] b. Contains a polypeptide sequence (preferably composed of) having at least 90% sequence identity with any one of SEQ ID NO:2, 10, 12, 14, 16, 18, 26, 34, 64 or 65.

[0496] 67. The polypeptide, method, or use according to any one of embodiments 24-26 or 49-66, wherein the polypeptide:

[0497] a. Encoded by a nucleotide sequence having at least 95% sequence identity with any one of SEQ ID NO: 1, 9, 11, 13, 15, 17, 25 or 33; or

[0498] b. Contains a polypeptide sequence (preferably composed of) having at least 95% sequence identity with any one of SEQ ID NO:2, 10, 12, 14, 16, 18, 26, 34, 64 or 65.

[0499] 68. The polypeptide, method, or use according to any one of embodiments 24-26 or 49-67, wherein the polypeptide:

[0500] a. Encoded by a nucleotide sequence having at least 99% sequence identity with any one of SEQ ID NO: 1, 9, 11, 13, 15, 17, 25 or 33; or

[0501] b. Contains a polypeptide sequence (preferably composed of) having at least 99% sequence identity with any one of SEQ ID NO:2, 10, 12, 14, 16, 18, 26, 34, 64 or 65.

[0502] 69. The polypeptide, method, or use according to any one of embodiments 24-26 or 49-68, wherein the polypeptide:

[0503] a. Encoded by a nucleotide sequence having at least 99.9% sequence identity with any one of SEQ ID NO: 1, 9, 11, 13, 15, 17, 25 or 33; or

[0504] b. Contains a polypeptide sequence (preferably composed of) having at least 99.9% sequence identity with any one of SEQ ID NO:2, 10, 12, 14, 16, 18, 26, 34, 64 or 65.

[0505] 70. The polypeptide, method, or use according to any of the preceding embodiments, wherein the polypeptide is applied at or near the site of injury, preferably wherein the polypeptide is applied intrathecally.

[0506] 71. The polypeptide, method, or use according to any of the foregoing embodiments, wherein the polypeptide does not further include a domain that binds to a cell receptor.

[0507] 72. The polypeptide, method, or use according to any of the foregoing embodiments, wherein the polypeptide lacks the functional H of clostridial neurotoxin. CIt has a structural domain and also lacks any functionally equivalent exogenous ligand targeting portion (TM).

[0508] 73. The polypeptide, method, or use according to any of the foregoing embodiments, wherein the polypeptide is not expressed in the subject's cells.

[0509] 74. The polypeptide, method, or use according to any of the preceding embodiments, wherein the clostridial sequence of the polypeptide consists of a clostridial neurotoxin light chain (L-chain) or a fragment thereof; and / or a fragment of a clostridial neurotoxin heavy chain (H-chain).

[0510] 75. The polypeptide, method, or use according to any of the preceding embodiments, wherein the polypeptide further comprises one or more non-clostridium neurotoxin sequences.

[0511] 76. The polypeptide, method, or use according to embodiment 75, wherein one or more non-clostridium neurotoxin sequences do not bind to cell receptors.

[0512] 77. The polypeptide, method, or use according to embodiment 75 or 76, wherein one or more nonclostridium neurotoxin sequences do not contain a ligand for a cell receptor.

[0513] 78. A polypeptide, method, or use according to any one of embodiments 1-40 or 49-77, wherein the polypeptide is a modified BoNT / A or a fragment thereof, comprising a modification at one or more amino acid residues selected from: ASN 886, ASN 905, GLN 915, ASN 918, GLU 920, ASN 930, ASN 954, SER 955, GLN 991, GLU 992, GLN 995, ASN 1006, ASN 1025, ASN 1026, ASN 1032, ASN 1043, ASN 1046, ASN 1052, ASP 1058, HIS 1064, ASN 1080, GLU 1081, GLU 1083, ASP 1086, ASN 1188, ASP 1213, GLY1215, ASN1216, GLN 1229, ASN 1242, ASN 1243, SER 1274, and THR 1277, where the modifiers are selected from:

[0514] vi. Replace surface-exposed acidic amino acid residues with basic amino acid residues;

[0515] vii. Replace surface-exposed acidic amino acid residues with uncharged amino acid residues;

[0516] viii. Replace uncharged surface-exposed amino acid residues with basic amino acid residues;

[0517] ix. Insertion of basic amino acid residues; and

[0518] x. Lacking surface-exposed acidic amino acid residues.

[0519] 79. The polypeptide, method, or use according to any one of embodiments 1-26 or 41-77, wherein the polypeptide comprises a BoNT / A light chain and a translocation domain and a BoNT / B receptor-binding domain (H). C The nesting of BoNTs (structural domains).

[0520] sequence list

[0521] The residue / codon is optional when the initial Met amino acid residue or the corresponding initial codon is indicated in any of the following SEQ ID NO.

[0522] SEQ ID NO:1 - Nucleotide sequence of recombinant non-catalytically active BoNT / A (rBoNT / A(0))

[0523] The polypeptide sequence of SEQ ID NO:2-rBoNT / A(0)

[0524] SEQ ID NO:3-rLH N / A nucleotide sequence (light chain plus translocation domain only).

[0525] SEQ ID NO:4-rLH N / A polypeptide sequence

[0526] Nucleotide sequence of SEQ ID NO:5-rL / A (light chain only)

[0527] The polypeptide sequence of SEQ ID NO:6-rL / A

[0528] SEQ ID NO:7-rH C / A nucleotide sequence

[0529] SEQ ID NO:8-rH C / A polypeptide sequence

[0530] Nucleotide sequence of SEQ ID NO:9-rBoNT / B(0)

[0531] The polypeptide sequence of SEQ ID NO:10-rBoNT / B(0)

[0532] Nucleotide sequence of SEQ ID NO:11-rBoNT / C(0)

[0533] The polypeptide sequence of SEQ ID NO:12-rBoNT / C(0)

[0534] Nucleotide sequence of SEQ ID NO:13-rBoNT / E(0)

[0535] The polypeptide sequence of SEQ ID NO:14-rBoNT / E(0)

[0536] Nucleotide sequence of SEQ ID NO:15-rBoNT / F(0)

[0537] The polypeptide sequence of SEQ ID NO:16-rBoNT / F(0)

[0538] The nucleotide sequence (His-tagged) of SEQ ID NO:17-rBoNT / A(0)

[0539] The polypeptide sequence (His-tagged) of SEQ ID NO:18-rBoNT / A(0)

[0540] SEQ ID NO:19-rLH N / A nucleotide sequence (His-tagged)

[0541] SEQ ID NO:20-rLH N / A polypeptide sequence (His-tagged)

[0542] SEQ ID NO:21-rH C / A nucleotide sequence (His-tagged)

[0543] SEQ ID NO:22-rH C / A polypeptide sequence (His-tagged)

[0544] The nucleotide sequence (His-tagged) of SEQ ID NO:23-rLc / A

[0545] The polypeptide sequence of SEQ ID NO:24-rLc / A (His-tagged)

[0546] Nucleotide sequence (His-tagged) of SEQ ID NO:25-rBoNT / FA(0)

[0547] The polypeptide sequence (His-tagged) of SEQ ID NO:26-rBoNT / FA(0)

[0548] SEQ ID NO:27-rLH N / FA nucleotide sequence (His-tagged)

[0549] SEQ ID NO:28-rLH N / FA polypeptide sequence (His-tagged)

[0550] SEQ ID NO:29-rH C / FA nucleotide sequence (His-tagged)

[0551] SEQ ID NO:30-rH C / FA polypeptide sequence (His-tagged)

[0552] Nucleotide sequence of SEQ ID NO:31-rLC / FA (His-tagged)

[0553] The polypeptide sequence of SEQ ID NO:32-rLC / FA (His-tagged)

[0554] The nucleotide sequence (His-tagged) of SEQ ID NO:33-rBoNT / F(0)

[0555] The polypeptide sequence (His-tagged) of SEQ ID NO:34-rBoNT / F(0)

[0556] SEQ ID NO:35-rL H N / F nucleotide sequence (His marker)

[0557] SEQ ID NO:36-rL H N / F polypeptide sequence (His-tagged)

[0558] SEQ ID NO:37-rH C / F nucleotide sequence (His-tagged)

[0559] SEQ ID NO:38-rH C / F polypeptide sequence (His-tagged)

[0560] Nucleotide sequence of SEQ ID NO:39-rLC / F (His-tagged)

[0561] The polypeptide sequence of SEQ ID NO:40-rLC / F (His-tagged)

[0562] SEQ ID NO:41 - rH of cations C / A nucleotide sequence (His-tagged)

[0563] SEQ ID NO:42 - rH of cations C / A polypeptide sequence (His-tagged)

[0564] SEQ ID NO:43-rH C The nucleotide sequence of / AB (His-tagged)

[0565] SEQ ID NO:44-rH C / AB polypeptide sequence (His-tagged)

[0566] SEQ ID NO:45-rH C The nucleotide sequence of variant Y1117V H1253K (His-tagged)

[0567] SEQ ID NO:46-rH C / A variant Y1117V H1253K polypeptide sequence (His-tagged)

[0568] SEQ ID NO:47-rH C The nucleotide sequence of variant A Y1117V F1252Y H1253K L1278F (His-tagged)

[0569] SEQ ID NO:48-rH C The polypeptide sequence of variant Y1117V F1252Y H1253K L1278F (His-tagged).

[0570] SEQ ID NO:49-rH C The nucleotide sequence of variant A Y1117V F1252Y H1253K L1278H (His-tagged)

[0571] SEQ ID NO:50-rH C The polypeptide sequence of variant Y1117V F1252Y H1253K L1278H (His-tagged)

[0572] SEQ ID NO:51-BoNT / A polypeptide sequence-Uniprot P10845

[0573] SEQ ID NO:52-BoNT / B polypeptide sequence-Uniprot P10844

[0574] SEQ ID NO:53-BoNT / C polypeptide sequence-Uniprot P18640

[0575] SEQ ID NO:54-BoNT / D polypeptide sequence-Uniprot P19321

[0576] SEQ ID NO:55-BoNT / E polypeptide sequence-Uniprot Q00496

[0577] SEQ ID NO:56-BoNT / F peptide sequence-Uniprot A7GBG3

[0578] SEQ ID NO:57-BoNT / G polypeptide sequence-Uniprot Q60393

[0579] SEQ ID NO:58 - Polypeptide sequence of TeNT - Uniprot P04958

[0580] The polypeptide sequence of SEQ ID NO:59-BoNT / X

[0581] The nucleotide sequence of SEQ ID NO:60-mrBoNT / A

[0582] The polypeptide sequence of SEQ ID NO:61-mrBoNT / A

[0583] SEQ ID NO:62 - Unmodified BoNT / A1 polypeptide sequence

[0584] The polypeptide sequence of SEQ ID NO:63-mrBoNT / AB

[0585] The polypeptide sequence of SEQ ID NO:64-mrBoNT / AB(0)

[0586] The polypeptide sequence of SEQ ID NO:65-MrBoNT / A(0)

[0587] Nucleotide sequence of SEQ ID NO:1-rBoNT / A(0)

[0588] ATGCCATTCGTCAACAAGCAATTCAACTACAAAGACCCAGTCAACGGCGTCGACAT

[0589] CGCATACATCAAGATTCCGAACGCCGGTCAAATGCAGCCGGTTAAGGCTTTTAAGA

[0590] TCCACAACAAGATTTGGGTTATCCCGGAGCGTGACACCTTCACGAACCCGGAAGAA

[0591] GGCGATCTGAACCCGCCACCGGAAGCGAAGCAAGTCCCTGTCAGCTACTACGATTC

[0592] GACGTACCTGAGCACGGATAACGAAAAAGATAACTACCTGAAAGGTGTGACCAAG

[0593] CTGTTCGAACGTATCTACAGCACGGATCTGGGTCGCATGCTGCTGACTAGCATTGTT

[0594] CGCGGTATCCCGTTCTGGGGTGGTAGCACGATTGACACCGAACTGAAGGTTATCGA

[0595] CACTAACTGCATTAACGTTATTCAACCGGATGGTAGCTATCGTAGCGAAGAGCTGA

[0596] ATCTGGTCATCATTGGCCCGAGCGCAGACATTATCCAATTCGAGTGCAAGAGCTTTG

[0597] GTCACGAGGTTCTGAATCTGACCCGCAATGGCTATGGTAGCACCCAGTACATTCGTT

[0598] TTTCGCCGGATTTTACCTTCGGCTTTGAAGAGAGCCTGGAGGTTGATACCAATCCGT

[0599] TGCTGGGTGCGGGCAAATTCGCTACCGATCCGGCTGTCACGCTGGCCCATcAACTGA

[0600] TCtACGCAGGCCACCGCCTGTACGGCATTGCCATCAACCCAAACCGTGTGTTCAAGG

[0601] TTAATACGAATGCATACTACGAGATGAGCGGCCTGGAAGTCAGCTTCGAAGAACTG

[0602] CGCACCTTCGGTGGCCATGACGCTAAATTCATTGACAGCTTGCAAGAGAATGAGTT

[0603] CCGTCTGTACTACTATAACAAATTCAAAGACATTGCAAGCACGTTGAACAAGGCCA

[0604] AAAGCATCGTTGGTACTACCGCGTCGTTGCAGTATATGAAGAATGTGTTTAAAGAG

[0605] AAGTACCTGCTGTCCGAGGATACCTCCGGCAAGTTTAGCGTTGATAAGCTGAAGTTT

[0606] GACAAACTGTACAAGATGCTGACCGAGATTTACACCGAGGACAACTTTGTGAAATT

[0607] CTTCAAAGTGTTGAATCGTAAAACCTATCTGAATTTTGACAAAGCGGTTTTCAAGAT

[0608] TAACATCGTGCCGAAGGTGAACTACACCATCTATGACGGTTTTAACCTGCGTAACAC

[0609] CAACCTGGCGGCGAACTTTAACGGTCAGAATACGGAAATCAACAACATGAATTTCA

[0610] CGAAGTTGAAGAACTTCACGGGTCTGTTCGAGTTCTATAAGCTGCTGTGCGTGCGCG

[0611] GTATCATCACCAGCAAAACCAAAAGCCTGGACAAAGGCTACAACAAGGCGCTGAA

[0612] TGACCTGTGCATTAAGGTAAACAATTGGGATCTGTTCTTTTCGCCATCCGAAGATAA

[0613] TTTTACCAACGACCTGAACAAGGGTGAAGAAATCACCAGCGATACGAATATTGAAG

[0614] CAGCGGAAGAGAATATCAGCCTGGATCTGATCCAGCAGTACTATCTGACCTTTAAC

[0615] TTCGACAATGAACCGGAGAACATTAGCATTGAGAATCTGAGCAGCGACATTATCGG

[0616] TCAGCTGGAACTGATGCCGAATATCGAACGTTTCCCGAACGGCAAAAAGTACGAGC

[0617] TGGACAAGTACACTATGTTCCATTACCTGCGTGCACAGGAGTTTGAACACGGTAAA

[0618] AGCCGTATCGCGCTGACCAACAGCGTTAACGAGGCCCTGCTGAACCCGAGCCGTGT

[0619] CTATACCTTCTTCAGCAGCGACTATGTTAAGAAAGTGAACAAAGCCACTGAGGCCG

[0620] CGATGTTCCTGGGCTGGGTGGAACAGCTGGTATATGACTTCACGGACGAGACGAGC

[0621] GAAGTGAGCACTACCGACAAAATTGCTGATATTACCATCATTATCCCGTATATTGGT

[0622] CCGGCACTGAACATTGGCAACATGCTGTACAAAGACGATTTTGTGGGTGCCCTGAT

[0623] CTTCTCCGGTGCCGTGATTCTGCTGGAGTTCATTCCGGAGATTGCGATCCCGGTGTT

[0624] GGGTACCTTCGCGCTGGTGTCCTACATCGCGAATAAGGTTCTGACGGTTCAGACCAT

[0625] CGATAACGCGCTGTCGAAACGTAATGAAAAATGGGACGAGGTTTACAAATACATTG

[0626] TTACGAATTGGCTGGCGAAAGTCAATACCCAGATCGACCTGATCCGTAAGAAAATG

[0627] AAAGAGGCGCTGGAGAATCAGGCGGAGGCCACCAAAGCAATTATCAACTACCAAT

[0628] ACAACCAGTACACGGAAGAAGAGAAGAATAACATTAACTTCAATATCGATGATTTG

[0629] AGCAGCAAGCTGAATGAATCTATCAACAAAGCGATGATCAATATCAACAAGTTTTT

[0630] GAATCAGTGTAGCGTTTCGTACCTGATGAATAGCATGATTCCGTATGGCGTCAAACG

[0631] TCTGGAGGACTTCGACGCCAGCCTGAAAGATGCGTTGCTGAAATACATTTACGACA

[0632] ATCGTGGTACGCTGATTGGCCAAGTTGACCGCTTGAAAGACAAAGTTAACAATACC

[0633] CTGAGCACCGACATCCCATTTCAACTGAGCAAGTATGTTGATAATCAACGTCTGTTG

[0634] AGCACTTTCACCGAGTATATCAAAAACATCATCAATACTAGCATTCTGAACCTGCGT

[0635] TACGAGAGCAATCATCTGATTGATCTGAGCCGTTATGCAAGCAAGATCAACATCGG

[0636] TAGCAAGGTCAATTTTGACCCGATCGATAAGAACCAGATCCAGCTGTTTAATCTGG

[0637] AATCGAGCAAAATTGAGGTTATCCTGAAAAACGCCATTGTCTACAACTCCATGTAC

[0638] GAGAATTTCTCCACCAGCTTCTGGATTCGCATCCCGAAATACTTCAACAGCATTAGC

[0639] CTGAACAACGAGTATACTATCATCAACTGTATGGAGAACAACAGCGGTTGGAAGGT

[0640] GTCTCTGAACTATGGTGAGATCATTTGGACCTTGCAGGACACCCAAGAGATCAAGC

[0641] AGCGCGTCGTGTTCAAGTACTCTCAAATGATCAACATTTCCGATTACATTAATCGTT

[0642] GGATCTTCGTGACCATTACGAATAACCGTCTGAATAACAGCAAGATTTACATCAAT

[0643] GGTCGCTTGATCGATCAGAAACCGATTAGCAACCTGGGTAATATCCACGCAAGCAA

[0644] CAACATTATGTTCAAATTGGACGGTTGCCGCGATACCCATCGTTATATCTGGATCAA

[0645] GTATTTCAACCTGTTTGATAAAGAACTGAATGAGAAGGAGATCAAAGATTTGTATG

[0646] ACAACCAATCTAACAGCGGCATTTTGAAGGACTTCTGGGGCGATTATCTGCAATAC

[0647] GATAAGCCGTACTATATGCTGAACCTGTATGATCCGAACAAATATGTGGATGTCAA

[0648] TAATGTGGGTATTCGTGGTTACATGTATTTGAAGGGTCCGCGTGGCAGCGTTATGAC

[0649] GACCAACATTTACCTGAACTCTAGCCTGTACCGTGGTACGAAATTCATCATTAAGAA

[0650] ATATGCCAGCGGCAACAAAGATAACATTGTGCGTAATAACGATCGTGTCTACATCA

[0651] ACGTGGTCGTGAAGAATAAAGAGTACCGTCTGGCGACCAACGCTTCGCAGGCGGGT

[0652] GTTGAGAAAATTCTGAGCGCGTTGGAGATCCCTGATGTCGGTAATCTGAGCCAAGT

[0653] CGTGGTTATGAAGAGCAAGAACGACCAGGGTATCACTAACAAGTGCAAGATGAAC

[0654] CTGCAAGACAACAATGGTAACGACATCGGCTTTATTGGTTTCCACCAGTTCAACAAT

[0655] ATTGCTAAACTGGTAGCGAGCAATTGGTACAATCGTCAGATTGAGCGCAGCAGCCG

[0656] TACTTTGGGCTGTAGCTGGGAGTTTATCCCGGTCGATGATGGTTGGGGCGAACGTCC

[0657] GCTG

[0658] The polypeptide sequence of SEQ ID NO:2-rBoNT / A(0)

[0659] MPFVNKQFNYKDPVNGVDIAYIKIPNAGQMQPVKAFKIHNKIWVIPERDTFTNPEEGDL

[0660] NPPPEAKQVPVSYYDSTYLSTDNEKDNYLKGVTKLFERIYSTDLGRMLLTSIVRGIPFW

[0661] GGSTIDTELKVIDTNCINVIQPDGSYRSEELNLVIIGPSADIIQFECKSFGHEVLNLTRNGY

[0662] GSTQYIRFSPDFTFGFEESLEVDTNPLLGAGKFATDPAVTLAHQLIYAGHRLYGIAINPNR

[0663] VFKVNTNAYYEMSGLEVSFEELRTFGGHDAKFIDSLQENEFRLYYYNKFKDIASTLNKA

[0664] KSIVGTTASLQYMKNVFKEKYLLSEDTSGKFSVDKLKFDKLYKMLTEIYTEDNFVKFFK

[0665] VLNRKTYLNFDKAVFKINIVPKVNYTIYDGFNLRNTNLAANFNGQNTEINNMNFTKLK

[0666] NFTGLFEFYKLLCVRGIITSKTKSLDKGYNKALNDLCIKVNNWDLFFSPSEDNFTNDLN

[0667] KGEEITSDTNIEAAEENISLDLIQQYYLTFNFDNEPENISIENLSSDIIGQLELMPNIERFPN

[0668] GKKYELDKYTMFHYLRAQEFEHGKSRIALTNSVNEALLNPSRVYTFFSSDYVKKVNKA

[0669] TEAAMFLGWVEQLVYDFTDETSEVSTTDKIADITIIIPYIGPALNIGNMLYKDDFVGALIF

[0670] SGAVILLEFIPEIAIPVLGTFALVSYIANKVLTVQTIDNALSKRNEKWDEVYKYIVTNWL

[0671] AKVNTQIDLIRKKMKEALENQAEATKAIINYQYNQYTEEEKNNINFNIDDLSSKLNESIN

[0672] KAMININKFLNQCSVSYLMNSMIPYGVKRLEDFDASLKDALLKYIYDNRGTLIGQVDRL

[0673] KDKVNNTLSTDIPFQLSKYVDNQRLLSTFTEYIKNIINTSILNLRYESNHLIDLSRYASKIN

[0674] IGSKVNFDPIDKNQIQLFNLESSKIEVILKNAIVYNSMYENFSTSFWIRIPKYFNSISLNNE

[0675] YTIINCMENNSGWKVSLNYGEIIWTLQDTQEIKQRVVFKYSQMINISDYINRWIFVTITN

[0676] NRLNNSKIYINGRLIDQKPISNLGNIHASNNIMFKLDGCRDTHRYIWIKYFNLFDKELNE

[0677] KEIKDLYDNQSNSGILKDFWGDYLQYDKPYYMLNLYDPNKYVDVNNVGIRGYMYLK

[0678] GPRGSVMTTNIYLNSSLYRGTKFIIKKYASGNKDNIVRNNDRVYINVVVKNKEYRLATN

[0679] ASQAGVEKILSALEIPDVGNLSQVVVMKSKNDQGITNKCKMNLQDNNGNDIGFIGFHQ

[0680] FNNIAKLVASNWYNRQIERSSRTLGCSWEFIPVDDGWGERPL

[0681] SEQ ID NO:3-rLH N / A nucleotide sequence

[0682] atggagttcgttaacaaacagttcaactataaagacccagttaacggtgttgacattgcttacatcaaaatcccgaacgctggccagatgcag

[0683] ccggtaaaggcattcaaaatccacaacaaaatctgggttatcccggaacgtgatacctttactaacccggaagaaggtgacctgaacccgc

[0684] caccggaagcgaaacaggtgccggtatcttactatgactccacctacctgtctaccgataacgaaaaggacaactacctgaaaggtgttact

[0685] aaactgttcgagcgtatttactccaccgacctgggccgtatgctgctgactagcatcgttcgcggtatcccgttctggggcggttctaccatcg

[0686] ataccgaactgaaagtaatcgacactaactgcatcaacgttattcagccggacggttcctatcgttccgaagaactgaacctggtgatcatcg

[0687] gcccgtctgctgatatcatccagttcgagtgtaagagctttggtcacgaagttctgaacctcacccgtaacggctacggttccactcagtacat

[0688] ccgtttctctccggacttcaccttcggttttgaagaatccctggaagtagacacgaacccactgctgggcgctggtaaattcgcaactgatcct

[0689] gcggttaccctggctcacgaactgattcatgcaggccaccgcctgtacggtatcgccatcaatccgaaccgtgtcttcaaagttaacaccaa

[0690] cgcgtattacgagatgtccggtctggaagttagcttcgaagaactgcgtacttttggcggtcacgacgctaaattcatcgactctctgcaaga

[0691] aaacgagttccgtctgtactactataacaagttcaaagatatcgcatccaccctgaacaaagcgaaatccatcgtgggtaccactgcttctctc

[0692] cagtacatgaagaacgtttttaaagaaaaatacctgctcagcgaagacacctccggcaaattctctgtagacaagttgaaattcgataaacttt

[0693] acaaaatgctgactgaaatttacaccgaagacaacttcgttaagttctttaaagttctgaaccgcaaaacctatctgaacttcgacaaggcagt

[0694] attcaaaatcaacatcgtgccgaaagttaactacactatctacgatggtttcaacctgcgtaacaccaacctggctgctaattttaacggccag

[0695] aacacggaaatcaacaacatgaacttcacaaaactgaaaaacttcactggtctgttcgagttttacaagctgctgtgcGTCGACGGC

[0696] ATCATTACCTCCAAAACTAAATCTGACGATGACGATAAAAACAAAGCGCTGAACCT

[0697] GCAGtgtatcaaggttaacaactgggatttattcttcagcccgagtgaagacaacttcaccaacgacctgaacaaaggtgaagaaatcac

[0698] ctcagatactaacatcgaagcagccgaagaaaacatctcgctggacctgatccagcagtactacctgacctttaatttcgacaacgagccg

[0699] gaaaacatttctatcgaaaacctgagctctgatatcatcggccagctggaactgatgccgaacatcgaacgtttcccaaacggtaaaaagta

[0700] cgagctggacaaatataccatgttccactacctgcgcgcgcaggaatttgaacacggcaaatcccgtatcgcactgactaactccgttaacg

[0701] aagctctgctcaacccgtcccgtgtatacaccttcttctctagcgactacgtgaaaaaggtcaacaaagcgactgaagctgcaatgttcttgg

[0702] gttgggttgaacagcttgtttatgattttaccgacgagacgtccgaagtatctactaccgacaaaattgcggatatcactatcatcatcccgtac

[0703] atcggtccggctctgaacattggcaacatgctgtacaaagacgacttcgttggcgcactgatcttctccggtgcggtgatcctgctggagttc

[0704] atcccggaaatcgccatcccggtactgggcacctttgctctggtttcttacattgcaaacaaggttctgactgtacaaaccatcgacaacgcg

[0705] ctgagcaaacgtaacgaaaaatgggatgaagtttacaaatatatcgtgaccaactggctggctaaggttaatactcagatcgacctcatccg

[0706] caaaaaaatgaaagaagcactggaaaaccaggcggaagctaccaaggcaatcattaactaccagtacaaccagtacaccgaggaagaa

[0707] aaaaacaacatcaacttcaacatcgacgatctgtcctctaaactgaacgaatccatcaacaaagctatgatcaacatcaacaagttcctgaac

[0708] cagtgctctgtaagctatctgatgaactccatgatcccgtacggtgttaaacgtctggaggacttcgatgcgtctctgaaagacgccctgctg

[0709] aaatacatttacgacaaccgtggcactctgatcggtcaggttgatcgtctgaaggacaaagtgaacaataccttatcgaccgacatcccttttc

[0710] agctcagtaaatatgtcgataaccaacgccttttgtccactctagaagcaCACCATCATCACcaccatcaccatcaccat

[0711] SEQ ID NO:4-rLH N / A polypeptide sequence

[0712] MEFVNKQFNYKDPVNGVDIAYIKIPNAGQMQPVKAFKIHNKIWVIPERDTFTNPEEGDL

[0713] NPPPEAKQVPVSYYDSTYLSTDNEKDNYLKGVTKLFERIYSTDLGRMLLTSIVRGIPFW

[0714] GGSTIDTELKVIDTNCINVIQPDGSYRSEELNLVIIGPSADIIQFECKSFGHEVLNLTRNGY

[0715] GSTQYIRFSPDFTFGFEESLEVDTNPLLGAGKFATDPAVTLAHELIHAGHRLYGIAINPNR

[0716] VFKVNTNAYYEMSGLEVSFEELRTFGGHDAKFIDSLQENEFRLYYYNKFKDIASTLNKA

[0717] KSIVGTTASLQYMKNVFKEKYLLSEDTSGKFSVDKLKFDKLYKMLTEIYTEDNFVKFFK

[0718] VLNRKTYLNFDKAVFKINIVPKVNYTIYDGFNLRNTNLAANFNGQNTEINNMNFTKLK

[0719] NFTGLFEFYKLLCVDGIITSKTKSDDDDKNKALNLQCIKVNNWDLFFSPSEDNFTNDLN

[0720] KGEEITSDTNIEAAEENISLDLIQQYYLTFNFDNEPENISIENLSSDIIGQLELMPNIERFPN

[0721] GKKYELDKYTMFHYLRAQEFEHGKSRIALTNSVNEALLNPSRVYTFFSSDYVKKVNKA

[0722] TEAAMFLGWVEQLVYDFTDETSEVSTTDKIADITIIIPYIGPALNIGNMLYKDDFVGALIF

[0723] SGAVILLEFIPEIAIPVLGTFALVSYIANKVLTVQTIDNALSKRNEKWDEVYKYIVTNWL

[0724] AKVNTQIDLIRKKMKEALENQAEATKAIINYQYNQYTEEEKNNINFNIDDLSSKLNESIN

[0725] KAMININKFLNQCSVSYLMNSMIPYGVKRLEDFDASLKDALLKYIYDNRGTLIGQVDRL

[0726] KDKVNNTLSTDIPFQLSKYVDNQRLLSTLEAHHHHHHHHHH Nucleotide sequence of SEQ ID NO:5-rL / A

[0727] ATGCCATTCGTCAACAAGCAATTCAACTACAAAGACCCAGTCAACGGCGTCGACAT

[0728] CGCATACATCAAGATTCCGAACGCCGGTCAAATGCAGCCGGTTAAGGCTTTTAAGA

[0729] TCCACAACAAGATTTGGGTTATCCCGGAGCGTGACACCTTCACGAACCCGGAAGAA

[0730] GGCGATCTGAACCCGCCACCGGAAGCGAAGCAAGTCCCTGTCAGCTACTACGATTC

[0731] GACGTACCTGAGCACGGATAACGAAAAAGATAACTACCTGAAAGGTGTGACCAAG

[0732] CTGTTCGAACGTATCTACAGCACGGATCTGGGTCGCATGCTGCTGACTAGCATTGTT

[0733] CGCGGTATCCCGTTCTGGGGTGGTAGCACGATTGACACCGAACTGAAGGTTATCGA

[0734] CACTAACTGCATTAACGTTATTCAACCGGATGGTAGCTATCGTAGCGAAGAGCTGA

[0735] ATCTGGTCATCATTGGCCCGAGCGCAGACATTATCCAATTCGAGTGCAAGAGCTTTG

[0736] GTCACGAGGTTCTGAATCTGACCCGCAATGGCTATGGTAGCACCCAGTACATTCGTT

[0737] TTTCGCCGGATTTTACCTTCGGCTTTGAAGAGAGCCTGGAGGTTGATACCAATCCGT

[0738] TGCTGGGTGCGGGCAAATTCGCTACCGATCCGGCTGTCACGCTGGCCCATGAACTG

[0739] ATCCACGCAGGCCACCGCCTGTACGGCATTGCCATCAACCCAAACCGTGTGTTCAA

[0740] GGTTAATACGAATGCATACTACGAGATGAGCGGCCTGGAAGTCAGCTTCGAAGAAC

[0741] TGCGCACCTTCGGTGGCCATGACGCTAAATTCATTGACAGCTTGCAAGAGAATGAG

[0742] TTCCGTCTGTACTACTATAACAAATTCAAAGACATTGCAAGCACGTTGAACAAGGC

[0743] CAAAAGCATCGTTGGTACTACCGCGTCGTTGCAGTATATGAAGAATGTGTTTAAAG

[0744] AGAAGTACCTGCTGTCCGAGGATACCTCCGGCAAGTTTAGCGTTGATAAGCTGAAG

[0745] TTTGACAAACTGTACAAGATGCTGACCGAGATTTACACCGAGGACAACTTTGTGAA

[0746] ATTCTTCAAAGTGTTGAATCGTAAAACCTATCTGAATTTTGACAAAGCGGTTTTCAA

[0747] GATTAACATCGTGCCGAAGGTGAACTACACCATCTATGACGGTTTTAACCTGCGTAA

[0748] CACCAACCTGGCGGCGAACTTTAACGGTCAGAATACGGAAATCAACAACATGAATT

[0749] TCACGAAGTTGAAGAACTTCACGGGTCTGTTCGAGTTCTATAAGCTGCTGggtctagaagc

[0750] aCACCATCATCACcaccatcaccatcaccat The polypeptide sequence of SEQ ID NO:6-rL / A

[0751] MPFVNKQFNYKDPVNGVDIAYIKIPNAGQMQPVKAFKIHNKIWVIPERDTFTNPEEGDL

[0752] NPPPEAKQVPVSYYDSTYLSTDNEKDNYLKGVTKLFERIYSTDLGRMLLTSIVRGIPFW

[0753] GGSTIDTELKVIDTNCINVIQPDGSYRSEELNLVIIGPSADIIQFECKSFGHEVLNLTRNGY

[0754] GSTQYIRFSPDFTFGFEESLEVDTNPLLGAGKFATDPAVTLAHELIHAGHRLYGIAINPNR

[0755] VFKVNTNAYYEMSGLEVSFEELRTFGGHDAKFIDSLQENEFRLYYYNKFKDIASTLNKA

[0756] KSIVGTTASLQYMKNVFKEKYLLSEDTSGKFSVDKLKFDKLYKMLTEIYTEDNFVKFFK

[0757] VLNRKTYLNFDKAVFKINIVPKVNYTIYDGFNLRNTNLAANFNGQNTEINNMNFTKLK

[0758] NFTGLFEFYKLLGLEAHHHHHHHHHH SEQ ID NO:7-rH C / A nucleotide sequence

[0759] ATGCATCATCACCATCACCACAAAAACATCATCAATACTAGCATTCTGAACCTGCGT

[0760] TACGAGAGCAATCATCTGATTGATCTGAGCCGTTATGCAAGCAAGATCAACATCGG

[0761] TAGCAAGGTCAATTTTGACCCGATCGATAAGAACCAGATCCAGCTGTTTAATCTGG

[0762] AATCGAGCAAAATTGAGGTTATCCTGAAAAACGCCATTGTCTACAACTCCATGTAC

[0763] GAGAATTTCTCCACCAGCTTCTGGATTCGCATCCCGAAATACTTCAACAGCATTAGC

[0764] CTGAACAACGAGTATACTATCATCAACTGTATGGAGAACAACAGCGGTTGGAAGGT

[0765] GTCTCTGAACTATGGTGAGATCATTTGGACCTTGCAGGACACCCAAGAGATCAAGC

[0766] AGCGCGTCGTGTTCAAGTACTCTCAAATGATCAACATTTCCGATTACATTAATCGTT

[0767] GGATCTTCGTGACCATTACGAATAACCGTCTGAATAACAGCAAGATTTACATCAAT

[0768] GGTCGCTTGATCGATCAGAAACCGATTAGCAACCTGGGTAATATCCACGCAAGCAA

[0769] CAACATTATGTTCAAATTGGACGGTTGCCGCGATACCCATCGTTATATCTGGATCAA

[0770] GTATTTCAACCTGTTTGATAAAGAACTGAATGAGAAGGAGATCAAAGATTTGTATG

[0771] ACAACCAATCTAACAGCGGCATTTTGAAGGACTTCTGGGGCGATTATCTGCAATAC

[0772] GATAAGCCGTACTATATGCTGAACCTGTATGATCCGAACAAATATGTGGATGTCAA

[0773] TAATGTGGGTATTCGTGGTTACATGTATTTGAAGGGTCCGCGTGGCAGCGTTATGAC

[0774] GACCAACATTTACCTGAACTCTAGCCTGTACCGTGGTACGAAATTCATCATTAAGAA

[0775] ATATGCCAGCGGCAACAAAGATAACATTGTGCGTAATAACGATCGTGTCTACATCA

[0776] ACGTGGTCGTGAAGAATAAAGAGTACCGTCTGGCGACCAACGCTTCGCAGGCGGGT

[0777] GTTGAGAAAATTCTGAGCGCGTTGGAGATCCCTGATGTCGGTAATCTGAGCCAAGT

[0778] CGTGGTTATGAAGAGCAAGAACGACCAGGGTATCACTAACAAGTGCAAGATGAAC

[0779] CTGCAAGACAACAATGGTAACGACATCGGCTTTATTGGTTTCCACCAGTTCAACAAT

[0780] ATTGCTAAACTGGTAGCGAGCAATTGGTACAATCGTCAGATTGAGCGCAGCAGCCG

[0781] TACTTTGGGCTGTAGCTGGGAGTTTATCCCGGTCGATGATGGTTGGGGCGAACGTCC

[0782] GCTG SEQ ID NO:8-rH C / A polypeptide sequence

[0783] MHHHHHHKNIINTSILNLRYESNHLIDLSRYASKINIGSKVNFDPIDKNQIQLFNLESSKIE

[0784] VILKNAIVYNSMYENFSTSFWIRIPKYFNSISLNNEYTIINCMENNSGWKVSLNYGEIIWT

[0785] LQDTQEIKQRVVFKYSQMINISDYINRWIFVTITNNRLNNSKIYINGRLIDQKPISNLGNIH

[0786] ASNNIMFKLDGCRDTHRYIWIKYFNLFDKELNEKEIKDLYDNQSNSGILKDFWGDYLQ

[0787] YDKPYYMLNLYDPNKYVDVNNVGIRGYMYLKGPRGSVMTTNIYLNSSLYRGTKFIIKK

[0788] YASGNKDNIVRNNDRVYINVVVKNKEYRLATNASQAGVEKILSALEIPDVGNLSQVVV

[0789] MKSKNDQGITNKCKMNLQDNNGNDIGFIGFHQFNNIAKLVASNWYNRQIERSSRTLGC

[0790] SWEFIPVDDGWGERPL

[0791] Nucleotide sequence of SEQ ID NO:9-rBoNT / B(0)

[0792] ATGCCGGTGACGATTAACAACTTCAACTACAACGACCCGATTGACAACAACAACAT

[0793] TATCATGATGGAACCGCCGTTTGCACGCGGCACGGGCCGTTATTACAAAGCGTTTA

[0794] AAATCACCGATCGTATTTGGATTATCCCGGAACGCTACACGTTTGGTTATAAACCGG

[0795] AAGACTTCAACAAAAGCTCTGGCATCTTCAACCGTGATGTTTGCGAATACTACGATC

[0796] CGGACTACCTGAACACCAACGATAAGAAAAACATTTTTCTGCAAACGATGATCAAA

[0797] CTGTTCAATCGCATTAAAAGCAAACCGCTGGGTGAAAAACTGCTGGAAATGATTAT

[0798] CAATGGCATTCCGTATCTGGGTGATCGTCGCGTGCCGCTGGAAGAATTTAACACCA

[0799] ATATCGCGAGTGTTACGGTCAACAAACTGATTTCCAATCCGGGTGAAGTCGAACGT

[0800] AAAAAAGGCATCTTCGCCAACCTGATCATCTTCGGCCCGGGTCCGGTGCTGAACGA

[0801] AAATGAAACCATTGATATCGGTATTCAGAACCATTTTGCCTCACGCGAAGGCTTCGG

[0802] CGGTATTATGCAAATGAAATTTTGCCCGGAATATGTGTCGGTTTTCAACAATGTTCA

[0803] GGAAAACAAAGGTGCAAGCATCTTTAATCGTCGCGGCTATTTCTCTGATCCGGCTCT

[0804] GATCCTGATGCACcAACTGATTtATGTGCTGCACGGCCTGTATGGTATCAAAGTGGAT

[0805] GACCTGCCGATCGTTCCGAACGAGAAAAAATTTTTCATGCAGAGCACCGACGCAAT

[0806] TCAAGCTGAAGAACTGTATACGTTTGGCGGTCAGGACCCGTCTATTATCACCCCGAG

[0807] CACCGACAAAAGCATCTACGATAAAGTGCTGCAAAACTTTCGTGGCATTGTTGACC

[0808] GCCTGAATAAAGTCCTGGTGTGTATCTCTGATCCGAACATCAACATCAACATCTACA

[0809] AAAACAAATTCAAAGACAAATACAAATTCGTTGAAGATTCTGAAGGCAAATATAGT

[0810] ATTGACGTCGAATCCTTTGATAAACTGTACAAAAGTCTGATGTTCGGTTTCACCGAA

[0811] ACGAACATCGCGGAAAACTACAAAATCAAAACCCGCGCCTCCTATTTCAGCGACTC

[0812] TCTGCCGCCGGTTAAAATCAAAAATCTGCTGGATAACGAAATTTATACGATCGAAG

[0813] AAGGTTTCAACATCAGCGATAAAGACATGGAAAAAGAATACCGTGGCCAGAATAA

[0814] AGCAATCAACAAACAGGCGTATGAAGAAATTAGTAAAGAACATCTGGCGGTCTACA

[0815] AAATTCAGATGTGCAAATCCGTGAAAGCCCCGGGTATTTGTATCGATGTTGACAAT

[0816] GAAGACCTGTTTTTCATCGCCGATAAAAACAGTTTTTCCGATGACCTGTCAAAAAAT

[0817] GAACGCATCGAATACAACACCCAATCGAACTACATCGAAAACGATTTCCCGATCAA

[0818] CGAACTGATTCTGGATACGGACCTGATTAGTAAAATCGAACTGCCGTCAGAAAACA

[0819] CCGAATCGCTGACGGACTTTAATGTTGATGTCCCGGTGTATGAAAAACAGCCGGCA

[0820] ATTAAGAAAATTTTTACCGATGAAAACACGATCTTCCAGTACCTGTACAGCCAAAC

[0821] CTTTCCGCTGGACATTCGCGATATCTCTCTGACGAGTTCCTTTGATGACGCACTGCT

[0822] GTTCAGCAACAAAGTGTACTCCTTTTTCTCAATGGATTACATCAAAACCGCTAACAA

[0823] AGTGGTTGAAGCGGGCCTGTTTGCCGGTTGGGTGAAACAGATCGTTAACGATTTCGT

[0824] CATCGAAGCCAACAAAAGTAACACGATGGATAAAATTGCTGATATCTCCCTGATTG

[0825] TCCCGTATATTGGCCTGGCACTGAATGTGGGTAACGAAACGGCGAAAGGCAATTTT

[0826] GAAAACGCCTTCGAAATTGCAGGCGCTTCAATCCTGCTGGAATTTATTCCGGAACTG

[0827] CTGATCCCGGTCGTGGGTGCGTTCCTGCTGGAATCTTACATCGACAACAAAAACAA

[0828] AATCATCAAAACCATTGATAACGCGCTGACGAAACGTAACGAAAAATGGTCAGATA

[0829] TGTACGGCCTGATTGTTGCCCAGTGGCTGAGCACCGTCAACACGCAATTTTACACCA

[0830] TCAAAGAAGGTATGTACAAAGCGCTGAATTATCAGGCGCAAGCCCTGGAAGAAATC

[0831] ATCAAATACCGCTACAACATCTACAGCGAAAAAGAAAAATCTAACATCAACATCGA

[0832] CTTTAATGATATCAACAGCAAACTGAACGAAGGTATCAACCAGGCAATCGATAACA

[0833] TCAACAACTTCATCAACGGCTGCTCAGTGTCGTATCTGATGAAGAAAATGATCCCGC

[0834] TGGCTGTTGAAAAACTGCTGGATTTTGACAACACCCTGAAGAAAAACCTGCTGAAC

[0835] TACATCGATGAAAACAAACTGTACCTGATCGGCTCAGCCGAATACGAAAAATCGAA

[0836] AGTGAACAAATACCTGAAAACCATCATGCCGTTTGACCTGAGTATTTACACCAACG

[0837] ATACGATCCTGATCGAAATGTTCAACAAATACAACTCCGAAATTCTGAACAATATT

[0838] ATCCTGAACCTGCGTTACAAAGACAACAATCTGATCGATCTGAGCGGCTATGGTGC

[0839] AAAAGTTGAAGTCTACGACGGTGTCGAACTGAACGATAAAAACCAGTTCAAACTGA

[0840] CCTCATCGGCTAACTCAAAAATTCGTGTGACGCAGAACCAAAACATCATCTTCAACT

[0841] CGGTCTTTCTGGACTTCAGCGTGTCTTTCTGGATTCGCATCCCGAAATATAAAAATG

[0842] ATGGCATCCAGAACTACATCCATAACGAATACACCATCATCAACTGTATGAAAAAC

[0843] AACAGTGGTTGGAAAATTTCCATCCGTGGCAACCGCATTATCTGGACCCTGATTGAT

[0844] ATCAATGGTAAACGAAAAGCGTGTTTTTCGAATACAACATCCGTGAAGATATCTC

[0845] TGAATACATCAATCGCTGGTTTTTCGTGACCATTACGAACAATCTGAACAATGCGAA

[0846] AATCTATATCAACGGCAAACTGGAAAGTAATACCGACATCAAAGATATTCGTGGAAG

[0847] TTATCGCCAACGGTGAAATCATCTTCAAACTGGATGGCGACATCGATCGCACCAG

[0848] TTCATTTGGATGAAATACTTCTCCATCTTCCAACACGGAACTGAGTCAGTCCAATATC

[0849] GAAGAACGCTACCAAAATCCAATCATACTCGGAATACCTGAAAGATTTCTGGGGTAA

[0850] CCCGCTGATGTACAACAAAGAATACTACATGTTCAACGCGGGCAACAAAAAACTCAT

[0851] ACATCAAACTGAAAAAAGATTCGCCGGTGGGTGAAATCCTGACCCGTAGCAAATAC

[0852] AACCAGAACTCTAAATACATCAACTATCGCGATCTGTACATTGGCGAAAAATTATTATT

[0853] ATCCGTCGCAAAAGCAACTCTCAGAGTATTAATGATGACATCGTGCGTAAAGAAAA

[0854] CTACATCTATCTGGATTTCTTTAATCTGAACCAAGAATGGCGCGTTTATACCTACAA

[0855] ATACTTCCAAAAAAAAAAAAGAAAACTGTTCCTGGCCCCGATTAGCGACAGCGATG

[0856] AATTTCAACACATCCAGATCAAAGAATACGATGAACAGCCGACGTATAGTTGC

[0857] CAACTGCTGTTCAAAAAAGACGAAGAATCCACCGATGAAATTGGCCTGATTGGTAT

[0858] CCACCGTTTCTATGAAAGCGGTATCGTTTTCGAAGAATACAAAGATTACTTCTGTAT

[0859] CTCTAAATGGTATCTGAAAAGTCAAACGCAAACCGTACAACCTGAAACTGGGCT

[0860] GCAACTGGCAATTTATCCCGAAAGACGAAGGCTGGACCGAA

[0861] The polypeptide sequence of SEQ ID NO:10-rBoNT / B(0)

[0862] MPVTINNFNYNDPIDNNNIIMMEPPFARGTGRYYKAFKITDRIWIIPERYTFGYKPEDFN

[0863] KSSGIFNRDVCEYYDPDYLNTNDKKNIFLQTMIKLFNRIKSKPLGEKLLEMIINGIPYLGD

[0864] RRVPLEEFNTNIASVTVNKLISNPGEVERKKGIFANLIIFGPGPVLNETIDIGIQNHFAS

[0865] REGFGGIMQMKFCPEYVSVFNNVQENKGASIFNRRGYFSDPALILMHQLIYVLHGLYGI

[0866] KVDDLPIVPNEKFFMQSTDAIQAEELYTFGGQDPSIITPSTDKSIYDKVLQNFRGIVDRL

[0867] NKVLVCISDPNINIYKNKFKDKYKFVEDSEGKYSIDVESFDKLYKSLMFGFTETNIAE

[0868] NYKIKTRASYFSDSLPPVKIKNLLDNEIYTIEEGFNISDKDMEKEYRGQNKAINKQAYEEI

[0869] SKEHLAVYKIQMCKSVKAPGICIDVDNEDLFFIADKNSFSDDLSKNERIEYNTQSNYIEN

[0870] DFPINELILDTDLISKIELPSENTESLTDFNVDVPVYEKQPAIKKIFTDENTIFQYLYSQTFP

[0871] LDIRDISLTSFDDALLFSNKVYSFFSMDYIKTANKVVEAGLFAGWVKQIVNDFVIEANK

[0872] SNTMDKIADISLIVPYIGLALNVGNETAKGNFENAFEIAGASILLEFIPELLIPVVGAFLLE

[0873] SYIDNKNKIIKTIDNALTKRNEKWSDMYGLIVAQWLSTVNTQFYTIKEGMYKALNYQA

[0874] QALEEIIKYRYNIYSEKEKSNIDNIFNDINSKLNEGINQAIDNINNFINGCSVSYLMKKMIP

[0875] LAVEKLLDFDNTLKKNLLNYIDENKLYLIGSAEEYEKSKVNKYLKTIMPFDLSIYTNDTILI

[0876] EMFNKYNSEILNNIILNLRYKDNNLIDLSGYGAKVEVYDGVELNDKNQFKLTSSANSKI

[0877] RVTQNQNIIFNSVFLDFSVSFWIRIPKYKNDGIQNYIHNEYTIINCMKNNSGWKISIRGNRI

[0878] IWTLIDINGKTKSVFFEYNIREDISEYINRWFFVTITNNLNNAKIYINGKLESNTDIKKDIRE

[0879] VIANGEIIFKLDGDIDRTQFIWMKYFSIFNTELSQSNIEERYKIQSYSEYLKDFWGNPLMY

[0880] NKEYYMFNAGNKNSYIKLKKDSPVGEILTRSKYNQNSKYINYRDLYIGEKFIIRRKSNSQ

[0881] SINDDIVRKEDYIYLDFFNLNQEWRVYTYKYFKKEEEKLFLAPISDSDEFYNTIQIKEYDE

[0882] QPTYSCQLLFKKDEESTDEIGLIGIHRFYESGIVFEEYKDYFCISKWYLKEVKRKPYNLKL

[0883] GCNWQFIPKDEGWTE

[0884] Nucleotide sequence of SEQ ID NO:11-rBoNT / C(0)

[0885] ATGCCGATCACGATTAATAATTTCAACTATAGCGATCCGGTGGACAATAAGAATAT

[0886] TCTGTATCTGGATACTCATCTGAATACGCTGGCTAACGAACCGGAGAAAGCGTTCC

[0887] GCATCACAGGCAACATCTGGGTTATTCCCGATCGCTTTTCACGCAACAGCAACCCTA

[0888] ATCTGAACAAACCTCCTCGTGTCACCAGTCCTAAATCCGGTTATTACGACCCAAACT

[0889] ATCTGAGTACGGATAGCGATAAAGATCCCTTTCTGAAAGAGATCATTAAGCTGTTC

[0890] AAACGCATTAACTCTCGCGAAATTGGGGAAGAGCTGATCTATCGGCTTTCGACAGA

[0891] TATCCCGTTCCCAGGTAACAATAATACCCCGATTAATACTTTCGACTTTGATGTTGA

[0892] TTTCAATTCTGTGGATGTGAAAACGCGTCAAGGCAATAATTGGGTGAAAACTGGTA

[0893] GCATTAACCCGAGTGTAATTATCACAGGTCCCCGTGAGAACATCATCGACCCGGAA

[0894] ACCTCTACCTTCAAGCTGACGAACAACACGTTTGCTGCACAGGAAGGGTTTGGTGC

[0895] CCTGTCAATCATTTCCATCTCACCGCGTTTCATGTTAACCTACTCCAATGCCACAAAT

[0896] GATGTTGGCGAAGGACGTTTTAGCAAATCAGAATTTTGCATGGACCCAATTCTCATT

[0897] CTGATGggCacGCTGAACaATGCGATGCACAACTTGTATGGCATTGCTATTCCAAACG

[0898] ATCAAACCATTAGCTCCGTTACCAGTAATATCTTCTATAGCCAGTATAATGTCAAAT

[0899] TGGAGTATGCCGAAATTTACGCCTTTGGAGGCCCGACCATTGACCTGATTCCGAAAT

[0900] CTGCACGCAAATACTTCGAAGAAAAGGCGTTAGATTACTATCGCAGCATCGCGAAA

[0901] CGCCTGAACTCGATTACCACGGCCAATCCGTCGTCGTTCAACAAATACATTGGTGAA

[0902] TATAAACAGAAACTGATTCGCAAATATCGGTTTGTCGTAGAAAGCTCTGGTGAAGT

[0903] GACTGTAAACCGCAACAAATTTGTCGAACTCTACAACGAGTTGACCCAAATCTTTAC

[0904] CGAGTTTAACTACGCAAAGATCTATAACGTACAGAACCGCAAGATTTATCTTAGCA

[0905] ATGTATACACACCGGTTACTGCGAACATCTTAGACGACAATGTGTATGATATTCAGA

[0906] ATGGCTTTAACATCCCGAAATCAAATCTGAACGTTCTGTTTATGGGCCAGAACCTGA

[0907] GTCGTAATCCAGCACTGCGTAAAGTGAACCCGGAAAATATGCTCTACTTGTTTACCA

[0908] AATTTTGCCACAAAGCGATTGATGGCCGCTCTCTCTATAACAAAACGCTGGATTGTC

[0909] GTGAGTTACTTGTGAAGAACACTGATTTACCGTTCATTGGGGATATCTCCGACGTGA

[0910] AAACCGATATCTTCCTGCGCAAAGACATTAATGAAGAAACGGAAGTCATCTATTAC

[0911] CCCGACAATGTGAGCGTTGATCAGGTCATTTTATCGAAGAACACCTCCGAACATGG

[0912] TCAGTTGGATTTGCTGTACCCTAGCATTGACTCGGAGAGTGAAATCCTTCCGGGCGA

[0913] AAATCAAGTGTTTTACGACAACCGTACCCAAAATGTTGATTATTTGAATTCTTATTA

[0914] CTACCTGGAATCTCAGAAATTGAGCGACAATGTGGAAGATTTCACGTTCACACGCT

[0915] CCATTGAGGAAGCGCTGGATAATAGCGCGAAAGTGTATACGTATTTCCCTACCTTG

[0916] GCGAATAAAGTAAATGCTGGTGTCCAGGGAGGCTTATTTCTGATGTGGGCGAATGA

[0917] TGTGGTAGAAGATTTTACGACCAATATTTTGCGTAAGGACACCTTAGATAAAATTAG

[0918] CGATGTTAGCGCCATCATCCCCTATATTGGCCCAGCACTGAATATCTCGAACTCTGT

[0919] GCGTCGCGGAAACTTCACCGAAGCATTTGCGGTGACCGGGGTTACTATTCTGTTGGA

[0920] AGCCTTTCCGGAGTTTACTATTCCGGCGCTGGGTGCGTTTGTGATTTATTCGAAAGT

[0921] ACAAGAACGCAATGAAATTATCAAAACCATCGATAATTGCCTGGAACAACGCATTA

[0922] AACGCTGGAAGGATTCTTATGAATGGATGATGGGCACCTGGTTATCCCGTATTATCA

[0923] CACAGTTTAACAACATCTCGTATCAGATGTACGATTCACTGAACTACCAAGCAGGG

[0924] GCGATCAAAGCCAAGATCGACTTAGAATACAAGAAATATTCAGGTAGCGATAAAG

[0925] AGAATATTAAAAGCCAGGTTGAAAACCTGAAGAACTCTCTGGATGTCAAAATTTCA

[0926] GAGGCTATGAACAACATTAACAAATTTATCCGCGAATGTAGCGTCACGTATCTGTTT

[0927] AAAAACATGCTCCCGAAAGTGATTGATGAGCTCAACGAGTTTGATCGCAACACAAA

[0928] GGCCAAACTGATTAACCTGATTGATAGTCACAATATTATTTTAGTCGGTGAAGTTGA

[0929] CAAGCTGAAGGCTAAGGTCAATAACAGCTTTCAGAACACTATTCCGTTTAATATTTT

[0930] CTCCTATACGAACAATAGTCTGCTGAAAGACATTATCAACGAATACTTCAACAATAT

[0931] TAATGACAGCAAAATTCTGAGCCTGCAGAATCGTAAGAATACGCTGGTAGATACCA

[0932] GTGGATATAATGCGGAAGTCTCAGAAGAGGGTGATGTACAGCTGAACCCGATCTTT

[0933] CCGTTCGACTTTAAACTGGGGTCTAGTGGTGAAGATCGCGGTAAAGTGATCGTTACC

[0934] CAAAACGAGAACATTGTGTATAACAGCATGTACGAGAGTTTCTCAATTTCTTTCTGG

[0935] ATTCGCATCAATAAATGGGTTTCTAATTTGCCTGGCTATACCATCATTGATAGCGTC

[0936] AAAAACAACTCGGGCTGGTCGATTGGCATTATTAGCAACTTTCTGGTGTTTACCCTG

[0937] AAACAGAATGAGGATTCGGAACAGAGCATTAACTTCTCCTACGACATCAGCAACAA

[0938] TGCACCAGGGTATAACAAATGGTTCTTCGTAACGGTGACGAACAATATGATGGGCA

[0939] ATATGAAAATCTACATTAACGGGAAACTTATCGACACCATTAAAGTGAAAGAGCTT

[0940] ACTGGGATCAATTTTAGTAAAACCATTACCTTTGAGATCAACAAAATTCCGGACAC

[0941] GGGTCTGATTACCTCCGATTCGGATAATATCAATATGTGGATTCGCGACTTTTATAT

[0942] CTTCGCCAAAGAACTTGATGGCAAAGATATCAACATTTTGTTTAATTCCCTGCAGTA

[0943] TACCAATGTCGTTAAGGACTATTGGGGCAATGATCTCCGCTACAATAAAGAATACT

[0944] ACATGGTTAACATCGACTATCTCAATCGCTACATGTATGCTAACTCGCGTCAAATTG

[0945] TGTTTAACACACGTCGTAACAACAACGATTTTAACGAAGGTTATAAAATCATTATCA

[0946] AACGGATCCGCGGCAATACGAACGATACTCGTGTTCGTGGCGGTGACATTCTGTATT

[0947] TCGACATGACGATTAATAATAAAGCGTACAATCTGTTCATGAAGAACGAAACCATG

[0948] TACGCCGATAACCATTCCACTGAAGATATCTACGCAATCGGACTTCGCGAACAGAC

[0949] CAAAGACATTAACGACAACATCATCTTTCAGATTCAACCGATGAATAATACCTACT

[0950] ACTATGCCTCCCAGATCTTCAAAAGTAATTTCAACGGCGAAAACATTTCAGGCATTT

[0951] GCTCAATCGGCACTTATCGGTTCCGGTTAGGTGGTGATTGGTATCGTCACAACTACC

[0952] TTGTTCCCACAGTGAAACAAGGCAACTATGCATCGCTCTTAGAAAGCACATCTACG

[0953] CATTGGGGTTTTGTGGCAGTCAGTGAA

[0954] The polypeptide sequence of SEQ ID NO:12-rBoNT / C(0)

[0955] MPITINNFNYSDPVDNKNILYLDTHLNTLANEPEKAFRITGNIWVIPDRFSRNSNPNLNKP

[0956] PRVTSPKSGYYDPNYLSTDSDKDPFLKEIIKLFKRINSREIGEELIYRLSTDIPFPGNNNTPI

[0957] NTFDFDVDFNSVDVKTRQGNNWVKTGSINPSVIITGPRENIIDPETSTFKLTNNTFAAQE

[0958] GFGALSIISISPRFMLTYSNATNDVGEGRFSKSEFCMDPILILMGTLNNAMHNLYGIAIPN

[0959] DQTISSVTSNIFYSQYNVKLEYAEIYAFGGPTIDLIPKSARKYFEEKALDYYRSIAKRLNSI

[0960] TTANPSSFNKYIGEYKQKLIRKYRFVVESSGEVTVNRNKFVELYNELTQIFTEFNYAKIY

[0961] NVQNRKIYLSNVYTPVTANILDDNVYDIQNGFNIPKSNLNVLFMGQNLSRNPALRKVNP

[0962] ENMLYLFTKFCHKAIDGRSLYNKTLDCRELLVKNTDLPFIGDISDVKTDIFLRKDINEET

[0963] EVIYYPDNVSVDQVILSKNTSEHGQLDLLYPSIDSESEILPGENQVFYDNRTQNVDYLNS

[0964] YYYLESQKLSDNVEDFFTTRSIEEALDNSAKVYTYFPTLANKVNAGVQGGLFLMWAND

[0965] VVEDFTTNILRKDTLDKISDVSAIIPYIGPALNISNSVRRGNFTEAFAVTGVTILLEAFPEF

[0966] TIPALGAFVIYSKVqCERNEIIKTIDNCLEQRIKRWKDSYEWMMGTWLSRIITQFNNISYQ

[0967] MYDSLNYQAGAIKAKIDLEYKKYSGSDKENIKSQVENLKNSLDVKISEAMNNINKFIRE

[0968] CSVTYLFKNMLPKVIDELNEFDRNTKAKLINLIDSHNIILVGEVDKLKAKVNNSFQNTIP

[0969] FNIFSYTNNSLLKDIINEYFNNINDSKILSLQNRKNTLVDTSGYNAEVSEEGDVQLNPIFPF

[0970] DFKLGSSGEDGRKVIVTQNENIVYNSMYESFSISFWIRINKWVSNLPGYTIIDSVKNNSG

[0971] WSIGIISNFLVFTLKQNEDSEQSINFSYDISNNAPGYNKWFFVTVTNNMMGNMKIYINGK

[0972] LIDTIKVKELTGINFSKTITFEINKIPDTGLITSDSDNINMWIRDFYIFAKELDGKDINILFNS

[0973] LQYTNVVKDYWGNDLRYNKEYYMVNIDYLNRYMYANSRQIVFNTRRNNNDFNEGYK

[0974] IIIKRIRGNTNDTRVRGGDILYFDMTINNKAYNLFMKNETMYADNHSTEDIYAIGLREQT

[0975] KDINDNIIFQIQPMNNTYYYASQIFKSNFNGENISGICSIGTYRFRLGGDWYRHNYLVPTV

[0976] KQGNYASLLESTSTHWGFVPVSE

[0977] Nucleotide sequence of SEQ ID NO:13-rBoNT / E(0)

[0978] atgccgaaaatcaactctttcaactacaacgacccggttaacgaccgtaccatcctgtatatcaaaccgggtggttgccaggagttctacaaa

[0979] tctttcaacatcatgaaaaacatctggatcatcccggaacgtaacgttatcggtaccaccccgcaggacttccacccgccgacctctctgaaa

[0980] aacggtgactcttcttactacgacccgaactacctccagtctgacgaagaaaaagaccgtttcctgaaaatcgttaccaaaatcttcaaccgt

[0981] atcaacaacaacctgtctggtggtatcctgctggaagaactgtctaaagctaacccgtacctgggtaacgacaacaccccggacaaccagt

[0982] tccacatcggtgacgcttctgctgttgaaatcaaattctctaacggttctcaggacatcctgctgccgaacgttatcatcatgggtgctgaacc

[0983] ggacctgttcgaaaccaactcttctaacatctctctgcgtaacaactacatgccgtctaaccacggtttcggttctatcgctatcgttaccttctct

[0984] ccggaatactctttccgtttcaacgacaacagcatgaacgagttcatccaggacccggctctgaccctgatgcaccaactgatctactctctg

[0985] cacggtctgtacggtgctaaaggtatcaccaccaaatacaccatcacccagaaacagaacccgctgatcaccaacatccgtggtaccaac

[0986] atcgaagagttcctgaccttcggtggtaccgacctgaacatcatcacctctgctcagtctaacgacatctacaccaacctgctggctgactac

[0987] aaaaaaatcgcttctaaactgtctaaagttcaggtttctaacccgctgctgaacccgtacaaagacgttttcgaagctaaatacggtctggaca

[0988] aagacgcttctggtatctactctgttaacatcaacaaattcaacgacatcttcaaaaaactgtactctttcaccgagttcgacctggcgaccaaa

[0989] ttccaggttaaatgccgtcagacctacatcggtcagtacaaatacttcaaactgtctaacctgctgaacgactctatctacaacatctctgaag

[0990] gttacaacatcaacaacctgaaagttaacttccgtggtcagaacgctaacctgaacccgcgtatcatcaccccgatcaccggtcgtggtctg

[0991] gttaaaaaaatcatccgtttctgcAAGAATATTGTAAGCGTTAAAGGAATAAGAAAAAGTATCtgcat

[0992] cgaaatcaacaacggtgaactgttcttcgttgcttctgaaaactcttacaacgacgacaacatcaacaccccgaaagaaatcgacgacacc

[0993] gttacctctaacaacaactacgaaaacgacctggaccaggttatcctgaacttcaactctgaatctgctccgggtctgtctgacgaaaaactg

[0994] aacctgaccatccagaacgacgcttacatcccgaaatacgactctaacggtacctctgacatcgaacagcacgacgttaacgaactgaacg

[0995] ttttcttctacctggacgctcagaaagttccggaaggtgaaaacaacgttaacctgacctcttctatcgacaccgctctgctggaacagccga

[0996] aaatctacaccttcttctcttctgagttcatcaacaacgttaacaaaccggttcaggctgctctgttcgtttcttggattcagcaggttctggttga

[0997] cttcaccaccgaagctaaccagaaatctaccgttgacaaaatcgctgacatctctatcgttgttccgtacatcggtctggctctgaacatcggt

[0998] aacgaagctcagaaaggtaacttcaaagacgctctggaactgctgggtgctggtatcctgctggagttcgaaccggaactgctgatcccga

[0999] ccatcctggttttcaccatcaaatctttcctgggttcttctgacaacaaaaacaaagttatcaaagctatcaacaacgctctgaaagaacgtgac

[1000] gaaaaatggaaagaagtttactctttcatcgtttctaactggatgaccaaaatcaacacccagttcaacaaacgtaaagaacagatgtaccag

[1001] gctctccagaaccaggttaacgctatcaaaaccatcatcgaatctaaatacaactcttacaccctggaagaaaaaaacgaactgaccaacaa

[1002] atacgacatcaaacagatcgaaaacgaactgaaccagaaagtttctatcgctatgaacaacatcgaccgtttcctgaccgaatcttctatctct

[1003] tacctgatgaaactcatcaacgaagttaaaatcaacaaactgcgtgaatacgacgaaaacgttaaaacctacctgctgaactacatcatcca

[1004] gcacggttctatcctgggtgaatctcagcaggaactgaactctatggttaccgacaccctgaacaactctatcccgttcaaactgtcttcttaca

[1005] ccgacgacaaaatcctGATCTCTTACTTCAACAAATTCTTTAAAcgcATTAAGAGTTCATCGGT

[1006] TctgaatATGCGGTACAAAAATGATAAAtatGTCGATACTTCTGGATATgatAGCAATATCA

[1007] ACATTAACGGCGACGTGTATAAATATccgACAAATAAAAACCAGTTTGGGATATATA

[1008] ACGACAAGctgTCGGAGGTCAATattTCTCAAAACGACtatATCattTACGATAATaaaTATA

[1009] AAAACTTTAGCATTAGTtttTGGGTTcgtATACCTAATtatGACAATaaaattGTAAATGTGAA

[1010] TAACGAGTATACCATTATAAACTGTATGcgcGACAATAACAGTGGTTGGAAGGTATC

[1011] GctgAACCATAATGAGATTATCTGGACCctgcagGATAATgcaGGTATAAACCAGAAACT

[1012] GGCTTTTAACTATGGAAACGCAAATGGGATCTCAGATTACATTaataaaTGGatttttGTTac

[1013] cATTACGAACGATcgcTTAGGCGACTCAAAACTTTATATTAATggcAATctgATAGATCA

[1014] GAAATCAATCTTAAATTTGGGCAATATTCATGTCTCTgatAACATCTTGTTCAAGATC

[1015] GTTAATTGCAGTTACACTcgtTATATTGGCATTCGTTACTTTAATATCTTCgataaaGAAct

[1016] gGACGAGACGGAAATCcagACTCTGTATTCAAACGAGCCCAATACTAATATATTGAA

[1017] AGATTTTTGGGGTAACTATCTTTTATATGATAAAGAATACTATCTCCTGaatGTATTGA

[1018] AGCCAAACAATTTCATAGATAGACGCAAGGATAGCACATTAAGTATCAACAATATC

[1019] AGATCTACTATActgttaGCAAATCGCCTcTACTCCggtATTAAAGTGAAGATTcagCGGGT

[1020] TAATAACTCCAGTACCAATGATAATCTGGTCCGTAAGAACGATCAGGTATACATCaat

[1021] TTCGTCGCGAGCAAAACTcatCTCTTCCCGCTTTACGCCgatACAGCTACGACAAACAA

[1022] GGAAAAAACCATAAAAATTTCCAGCTCCGGAAACAGATTCAATCAAGTAGTTGTAA

[1023] TGAACTCTGTGGGTaatAATTGTACGATGAACTTTaagAATAACAATGGGAACAATattG

[1024] GACTTTTGGGCTTcAAAGCCGACACAGTGGTGGCGTCCACCTGGTATTACACGcacAT

[1025] GcggGACCATACGAATTCGAACGGTTGCTTCTGGAACTTTATCTCGGAAgaaCACGGG

[1026] TGGCAAGAAAAA

[1027] The polypeptide sequence of SEQ ID NO:14-rBoNT / E(0)

[1028] MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTTPQDFHPPTSL

[1029] KNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGGILLEELSKANPYLGNDNTPD

[1030] NQFHIGDASAVEIKFSNGSQDILLPNVIIMGAEPDLFETNSSNISLRNNYMPSNHGFGSIAI

[1031] VTFSPEYSFRFNDNSMNEFIQDPALTLMHQLIYSLHGLYGAKGITTKYTITQKQNPLITNI

[1032] RGTNIEEFLTFGGTDLNIITSAQSNDIYTNLLADYKKIASKLSKVQVSNPLLNPYKDVFEA

[1033] KYGLDKDASGIYSVNINKFNDIFKKLYSFTEFDLATKFQVKCRQTYIGQYKYFKLSNLL

[1034] NDSIYNISEGYNINNLKVNFRGQNANLNPRIITPITGRGLVKKIIRFCKNIVSVKGIRKSICI

[1035] EINGLEFFVASENSYNDDNINTPKEIDDTVTSNNNYENDLDQVILNFNSESAPGLSDEK

[1036] LNLTIQNDAYIPKYDSNGTSDIEQHDVNELNVFFYLDAQKVPEGENNVNLTSSIDTALLE

[1037] QPKIYTFFSSEFINNVNKPVQAALFVSWIQQVLVDFTTEANQKSTVDKIADISIVVPYIGL

[1038] ALNIGNEAQKGNFKDALELLGAGILLEFEPELLIPTILVFTIKSFLGSSDNKNKVIKAINNA

[1039] LKERDEKWKEVYSFIVSNWMTKINTQFNKRKEQMYQALQNQVNAIKTIIESKYNSYTL

[1040] EEKNELTNKYDIKQIENELNQKVSIAMNNIDRFLTESSISYLMKLINEVKINKLREYDEN

[1041] VKTYLLNYIIQHGSILGESQQELNSMVTDTLNNSIPFKLSSYTDDKILISYFNKFFKRIKSS

[1042] SVLNMRYKNDKYVDTSGYDSNININGDVYKYPTNKNQFGIYNDKLSEVNISQNDYIIYD

[1043] NKYKNFSISFWVRIPNYDNKIVNVNNEYTIINCMRDNNSGWKVSLNHNEIIWTLQDNAG

[1044] INQKLAFNYGNANGISDYINKWIFVTITNDRLGDSKLYINGNLIDQKSILNLGNIHVSDNI

[1045] LFKIVNCSYTRYIGIRIFNIFDKELDETEIQTLYSNEPNTNILKDFWGNYLLYDKEYYLLN

[1046] VLKPNNFIDRRKDSTLSINNIRSTILLANRLYSGIKVKIQRVNNSSTNDNLVRKNDQVYIN

[1047] FVASKTHLFPLYADTATTNKEKTIKISSSGNRFNQVVVMNSVGNNCTMNFKNNNGNNI

[1048] GLLGFKADTVVASTWYYTHMRDHTNSNGCFWNFISEEHGWQEK

[1049] Nucleotide sequence of SEQ ID NO:15-rBoNT / F(0)

[1050] ATGCCGGTGGTCATCAACAGCTTCAACTACAACGACCCAGTAAACGACGACACGAT

[1051] CCTGTATATGCAAATCCCGTATGAAGAGAAGAGCAAGAAGTACTATAAGGCCTTTG

[1052] AAATCATGCGCAATGTGTGGATTATTCCGGAGCGTAATACGATTGGTACTGACCCA

[1053] AGCGACTTCGATCCACCTGCGTCTTTGGAAAACGGCTCGTCCGCATATTACGACCCG

[1054] AATTACCTGACCACCGATGCGGAGAAAGATCGTTATTTGAAAACCACCATCAAGCT

[1055] GTTCAAACGCATTAACAGCAATCCGGCAGGTGAGGTCCTGCTGCAAGAGATTAGCT

[1056] ACGCAAAGCCTTATCTGGGTAATGAGCATACGCCTATTAACGAGTTTCACCCGGTTA

[1057] CCCGCACTACCAGCGTTAACATCAAGTCCTCGACCAACGTGAAGTCTAGCATTATCC

[1058] TGAACCTGCTGGTTCTGGGTGCCGGTCCGGACATCTTCGAAAACTCTAGCTACCCGG

[1059] TGCGTAAACTGATGGATAGCGGCGGTGTTTATGACCCGAGCAATGACGGTTTTGGC

[1060] AGCATCAATATCGTGACGTTTAGCCCGGAGTACGAGTACACCTTCAATGATATCAG

[1061] CGGTGGTTACAATTCTTCTACCGAGAGCTTCATCGCCGACCCGGCGATCAGCCTGGC

[1062] ACACCAACTGATCTATGCATTGCATGGCTTGTACGGTGCCCGTGGTGTGACGTATAA

[1063] AGAGACTATCAAGGTTAAGCAGGCACCTCTGATGATTGCGGAAAAGCCGATTCGCC

[1064] TGGAAGAGTTCCTGACCTTCGGCGGTCAAGATTTGAACATCATTACCTCGGCCATGA

[1065] AAGAGAAAATCTATAACAATTTGCTGGCCAACTATGAAAAGATTGCAACGCGCTTG

[1066] TCTCGTGTTAACTCCGCTCCGCCGGAATACGACATTAATGAGTACAAAGACTACTTT

[1067] CAATGGAAATATGGCCTGGACAAAAATGCGGATGGTTCTTATACCGTGAATGAAAA

[1068] CAAATTCAATGAAATCTACAAGAAACTGTACAGCTTCACCGAAATCGATCTGGCGA

[1069] ACAAGTTCAAAGTCAAATGTCGTAATACCTACTTCATCAAATATGGCTTCCTGAAAG

[1070] TCCCGAACCTGCTGGACGATGACATCTATACCGTCAGCGAAGGCTTCAACATCGGC

[1071] AATCTGGCCGTGAATAATCGTGGTCAGAACATCAAACTGAATCCGAAAATCATTGA

[1072] CTCCATCCCAGACAAGGGCCTGGTTGAGAAAATCGTGAAGTTCTGCAAAAGCGTTA

[1073] TTCCGCGTAAAGGTACGAAAGCACCGCCTCGCCTGTGCATTCGCGTTAACAACCGT

[1074] GAGTTGTTCTTTGTGGCATCTGAAAGCAGCTACAACGAGAACGACATCAACACCCC

[1075] TAAAGAAATTGATGATACCACGAACCTGAATAACAATTATCGCAACAATCTGGACG

[1076] AGGTGATCCTGGATTACAATTCGGAAACCATTCCGCAAATTAGCAATCAGACGCTG

[1077] AACACCCTGGTTCAGGACGATAGCTACGTTCCGCGTTACGACTCCAATGGTACTAGC

[1078] GAGATTGAAGAACACAACGTAGTGGACTTGAACGTTTTCTTTTATCTGCACGCCCAG

[1079] AAGGTTCCGGAGGGCGAAACCAATATTAGCCTGACCAGCTCGATCGACACCGCGCT

[1080] GTCTGAGGAGAGCCAAGTCTACACCTTTTTCAGCAGCGAGTTTATCAACACTATTAA

[1081] CAAGCCAGTTCATGCTGCATTGTTTATCTCTTGGATTAACCAGGTGATTCGCGACTT

[1082] TACGACGGAGGCGACCCAGAAGTCTACCTTCGACAAAATTGCAGACATCTCCCTGG

[1083] TCGTCCCATACGTCGGCCTGGCGTTGAATATTGGCAATGAAGTTCAAAAAGAGAAC

[1084] TTCAAAGAAGCGTTCGAGCTGCTGGGTGCAGGCATCCTGCTGGAGTTCGTGCCGGA

[1085] ACTGTTGATCCCGACCATCCTGGTGTTCACCATTAAGAGCTTCATTGGATCCTCCGA

[1086] GAATAAGAACAAGATCATCAAGGCGATCAATAACAGCCTGATGGAGCGTGAAACG

[1087] AAGTGGAAAGAAATCTATAGCTGGATTGTTAGCAATTGGCTGACTCGTATTAACAC

[1088] GCAATTCAACAAGCGTAAAGAGCAAATGTACCAAGCCCTGCAAAACCAAGTTGACG

[1089] CCATCAAAACGGTAATTGAATACAAGTACAACAATTACACGAGCGATGAGCGCAAC

[1090] CGCCTGGAAAGCGAATACAACATCAACAACATTCGCGAAGAATTGAACAAGAAAG

[1091] TGAGCCTGGCGATGGAGAACATTGAGCGTTTTATCACCGAAAGCAGCATCTTTTACC

[1092] TGATGAAATTGATTAATGAGGCGAAAGTCTCGAAACTGCGTGAGTACGACGAAGGT

[1093] GTGAAAGAGTATCTGCTGGATTACATTAGCGAGCACCGTAGCATCTTGGGTAACTC

[1094] GGTTCAGGAGCTGAACGATCTGGTGACCTCTACCCTGAACAATAGCATCCCGTTCG

[1095] AACTGAGCAGCTATACCAATGACAAGATTCTGATTCTGTATTTCAATAAACTGTATA

[1096] AGAAGATCAAGGATAACAGCATTCTGGATATGCGTTACGAAAACAATAAGTTTATC

[1097] GACATTTCTGGTTACGGCAGCAACATTTCCATCAATGGCGATGTCTACATCTACAGC

[1098] ACCAATCGCAACCAGTTCGGCATCTACTCTAGCAAACCGAGCGAAGTTAACATCGC

[1099] ACAGAACAATGATATTATTTATAACGGTCGTTATCAAAACTTCTCTATCAGCTTTTG

[1100] GGTCCGTATCCCGAAGTACTTCAATAAAGTCAATCTGAATAATGAATACACGATCA

[1101] TCGACTGCATTCGCAATAACAACAGCGGTTGGAAAATCAGCCTGAATTACAACAAA

[1102] ATTATTTGGACCCTGCAAGATACGGCGGGTAACAATCAGAAACTGGTGTTTAACTA

[1103] CACGCAAATGATCAGCATTTCTGACTATATCAACAAGTGGATCTTTGTTACCATCAC

[1104] CAATAATCGTCTGGGCAATAGCCGTATTTACATCAACGGTAACCTGATTGATGAGA

[1105] AAAGCATCAGCAACCTGGGCGATATTCACGTCAGCGACAACATTCTGTTCAAAATT

[1106] GTTGGTTGTAACGATACCCGTTACGTCGGCATCCGTTATTTCAAGGTTTTCGATACG

[1107] GAGCTGGGTAAAACGGAAATCGAAACGTTGTACTCCGATGAACCAGATCCGAGCAT

[1108] TCTGAAGGACTTTTGGGGTAACTACTTGCTGTACAATAAACGTTACTATCTGCTGAA

[1109] TCTGTTGCGCACCGACAAGAGCATTACCCAAAACAGCAATTTCCTGAACATTAATC

[1110] AGCAACGCGGCGTATACCAAAAACCGAACATCTTCAGCAATACGCGCCTGTATACT

[1111] GGTGTTGAAGTGATCATTCGTAAGAACGGTAGCACCGACATTAGCAACACGGACAA

[1112] TTTCGTCCGTAAGAATGACCTGGCGTACATTAACGTCGTGGACCGTGATGTCGAGTA

[1113] TCGTCTGTACGCAGACATCAGCATTGCGAAACCGGAAAAGATTATCAAGCTGATCC

[1114] GTACCAGCAACAGCAACAACAGCCTGGGTCAGATCATTGTGATGGACAGCATTGGT

[1115] AATAACTGCACGATGAACTTCCAGAACAACAATGGTGGTAATATCGGTCTGCTGGG

[1116] TTTTCACAGCAATAATCTGGTTGCTTCCAGCTGGTACTACAATAACATTCGTAAAAA

[1117] CACGTCTAGCAATGGTTGTTTTTGGAGCTTTATCAGCAAAGAGCACGGCTGGCAAG

[1118] AAAAT

[1119] The polypeptide sequence of SEQ ID NO:16-rBoNT / F(0)

[1120] MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPERNTIGTDPSDFD

[1121] PPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRINSNPAGEVLLQEISYAKPYLGN

[1122] EHTPINEFHPPVTRTTSVNIKSSTVNKSSIILNLLVLGAGPDIHENSSYPVRKLMDSGGVYD

[1123] PSNDGFGSINIVTFSPEYETFNDISGGYNSSTESFIADPAISLAHQLIYALHGLYGARGVT

[1124] YKETIKVKQAPLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV

[1125] NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFTEIDLANKFKV

[1126] KCRNTYFIKYGFLKVPNLLDDDIYTVSEGFNIGNLAVNNRGQNIKLNPKIIDSIPDKGLVE

[1127] KIVKFCKSVIPRKGTKAPPRLCIRVNNRELFFVASESSYNDINTKEIDDTTTNLNNYR

[1128] NNLDEVILDYNSETIPQISNQTLNTLVQDDSYVPRYDSNGTSEIEEHNVVDLNVFFYLHA

[1129] QKVPEGETNISLTSSIDTALSEESQVYTFFSSEFINTINKPVHAALFISWINQVIRDFTTEAT

[1130] QKSTFDKIADISLVVPYVGLALNIGNEVQKENFKEAFELLGAGILLEFVPELLIPTILVFTI

[1131] KSFIGSSENKNKIIKAINNSLMERETKWKEIYSWIVSNWLTRINTQFNKRKEQMYQALQ

[1132] NQVDAIKTVIEYKYNNYTSDERNRLESEYNINNIREELNKKVSLAMENIERFITESSIFYL

[1133] MKLINEAKVSKLREYDEGVKEYLLDYISEHRSILGNSVQELNDLVTSTLNNSIPFELSSYT

[1134] NDKILILYFNKLYKKIKDNSILDMRYENNKFIDISGYGSNISINGDVYIYSTNRNQFGIYSS

[1135] KPSEVNIAQNNDIIYNGRYQNFSISFWVRIPKYFNKVNLNNEYTIIDCIRNNNSGWKISLN

[1136] YNKIIWTLQDTAGNNQKLVFNYTQMISISDYINKWIFVTITNNRLGNSRIYINGNLIDEKS

[1137] ISNLGDIHVSDNILFKIVGCNDTRYVGIRYFKVFDTELGKTEIETLYSDEPDPSILKDFWG

[1138] NYLLYNKRYYLLNLLRTDKSITQNSNFLNINQQRGVYQKPNIFSNTRLYTGVEVIIRKNG

[1139] STDISNTDNFVRKNDLAYINVVDRDVEYRLYADISIAKPEKIIKLIRTSNSNNSLGQIIVM

[1140] DSIGNNCTMNFQNNNGGNIGLLGFHSNNLVASSWYYNNIRKNTSSNGCFWSFISKEHG

[1141] WQEN

[1142] Nucleotide sequence (His-tag) of SEQ ID NO:17-rBoNT / A(0)

[1143] ATGCCGTTTGTGAACAAGCAGTTCAACTATAAAGATCCGGTTAATGGTGTGGATATC

[1144] GCCTATATCAAAATTCCGAATGCAGGTCAGATGCAGCCGGTTAAAGCCTTTAAAAT

[1145] CCATAACAAAATTTGGGTGATTCCGGAACGTGATACCTTTACCAATCCGGAAGAAG

[1146] GTGATCTGAATCCGCCTCCGGAAGCAAAACAGGTTCCGGTTAGCTATTATGATAGC

[1147] ACCTATCTGAGCACCGATAACGAGAAAGATAACTATCTGAAAGGTGTGACCAAACT

[1148] GTTTGAACGCATTTATAGTACCGATCTGGGTCGTATGCTGCTGACCAGCATTGTTCG

[1149] TGGTATTCCGTTTTGGGGTGGTAGCACCATTGATACCGAACTGAAAGTTATTGACAC

[1150] CAACTGCATTAATGTGATTCAGCCGGATGGTAGCTATCGTAGCGAAGAACTGAATC

[1151] TGGTTATTATTGGTCCGAGCGCAGATATCATTCAGTTTGAATGTAAAAGCTTTGGCC

[1152] ACGAAGTTCTGAATCTGACCCGTAATGGTTATGGTAGTACCCAGTATATTCGTTTCA

[1153] GTCCGGATTTTACCTTTGGCTTTGAAGAAAGCCTGGAAGTTGATACAAATCCGCTGT

[1154] TAGGTGCAGGTAAATTTGCAACCGATCCGGCAGTTACCCTGGCACACCAGCTGATTT

[1155] ATGCCGGTCATCGTCTGTATGGTATTGCCATTAATCCGAATCGTGTGTTCAAAGTGA

[1156] ATACCAACGCCTATTATGAAATGAGCGGTCTGGAAGTGAGTTTTGAAGAACTGCGT

[1157] ACCTTTGGTGGTCATGATGCCAAATTTATCGATAGCCTGCAAGAAAATGAATTTCGC

[1158] CTGTACTACTATAACAAATTCAAGGATATTGCGAGCACCCTGAATAAAGCCAAAAG

[1159] CATTGTTGGCACCACCGCAAGCCTGCAGTATATGAAAAATGTGTTTAAAGAAAAAT

[1160] ATCTGCTGAGCGAAGATACCAGCGGTAAATTTAGCGTTGACAAACTGAAATTCGAT

[1161] AAACTGTACAAGATGCTGACCGAGATTTATACCGAAGATAACTTCGTGAAGTTTTTC

[1162] AAAGTGCTGAACCGCAAAACCTACCTGAACTTTGATAAAGCCGTGTTCAAAATCAA

[1163] CATCGTGCCGAAAGTGAACTATACCATCTATGATGGTTTTAACCTGCGCAATACCAA

[1164] TCTGGCAGCAAACTTTAATGGTCAGAACACCGAAATCAACAACATGAACTTTACCA

[1165] AACTGAAGAACTTCACCGGTCTGTTCGAATTTTACAAACTGCTGTGTGTCGTGGCA

[1166] TTATTACCAGCAAAACCAAGGTCTGGATAAAGGCTACAATAAAGCCCTGAATGAT

[1167] CTGTGCATTAAGGTGAATAATTGGGACCTGTTTTTTAGCCCGAGCGAGGATAATTTC

[1168] ACCAACGATCTGAACAAAGGCGAAGAAATTACCAGCGATACCAATATTGAAGCAG

[1169] CCGAAGAAAACATTAGCCTGGATCTGATTCAGCAGTATTATCTGACCTTCAACTTCG

[1170] ATAATGAGCCGGAAAATATCAGCATTGAAAACCTGAGCAGCGATATTATTGGCCAG

[1171] CTGGAACTGATGCCGAATATTGAACGTTTTCCGAACGGCAAAAAATACGAGCTGGA

[1172] TAAATACACCATGTTCCATTATCTGCGTGCCCAAGAATTTGAACATGGTAAAAGCCG

[1173] TATTGCACTGACCAATAGCGTTAATGAAGCACTGCTGAACCCGAGCCGTGTTTATAC

[1174] CTTTTTTAGCAGCGATTACGTGAAAAAGGTTAACAAAGCAACCGAAGCAGCCATGT

[1175] TTTTAGGTTGGGTTGAACAGCTGGTTTATGATTTCACCGATGAAACCAGCGAAGTTA

[1176] GCACCACCGATAAAATTGCAGATATTACCATCATCATCCCGTATATCGGTCCGGCAC

[1177] TGAATATTGGCAATATGCTGTATAAAGACGATTTTGTGGGTGCCCTGATTTTTAGCG

[1178] GTGCAGTTATTCTGCTGGAATTTATTCCGGAAATTGCCATTCCGGTTCTGGGCACCT

[1179] TTGCACTGGTGAGCTATATTGCAAATAAAGTTCTGACCGTGCAGACCATCGATAATG

[1180] CACTGAGCAAACGTAACGAAAAATGGGATGAAGTGTACAAGTATATCGTGACCAAT

[1181] TGGCTGGCAAAAGTTAACACCCAGATTGACCTGATTCGCAAGAAGATGAAAGAAGC

[1182] ACTGGAAAATCAGGCAGAAGCAACCAAAGCCATTATCAACTATCAGTATAACCAGT

[1183] ACACCGAAGAAGAGAAAAATAACATCAACTTCAACATCGACGATCTGTCCAGCAAA

[1184] CTGAACGAAAGCATCAACAAAGCCATGATTAACATTAACAAATTTCTGAACCAGTG

[1185] CAGCGTGAGCTATCTGATGAATAGCATGATTCCGTATGGTGTGAAACGTCTGGAAG

[1186] ATTTTGATGCAAGCCTGAAAGATGCCCTGCTGAAATATATCTATGATAATCGTGGCA

[1187] CCCTGATTGGTCAGGTTGATCGTCTGAAAGATAAAGTGAACAACACCCTGAGTACC

[1188] GATATTCCTTTTCAGCTGAGCAAATATGTGGATAATCAGCGTCTGCTGTCAACCTTT

[1189] ACCGAATACATTAAGAACATCATCAACACCAGCATTCTGAACCTGCGTTATGAAAG

[1190] CAATCATCTGATTGATCTGAGCCGTTATGCCAGCAAAATCAATATAGGCAGCAAGG

[1191] TTAACTTCGACCCGATTGACAAAAATCAGATACAGCTGTTTAATCTGGAAAGCAGC

[1192] AAAATTGAGGTGATCCTGAAAAACGCCATTGTGTATAATAGCATGTACGAGAATTT

[1193] CTCGACCAGCTTTTGGATTCGTATCCCGAAATACTTTAATAGCATCAGCCTGAACAA

[1194] CGAGTACACCATTATTAACTGCATGGAAAACAATAGCGGCTGGAAAGTTAGCCTGA

[1195] ATTATGGCGAAATTATCTGGACCCTGCAGGATACCCAAGAAATCAAACAGCGTGTG

[1196] GTTTTCAAATACAGCCAGATGATTAATATCAGCGACTATATCAACCGCTGGATTTTT

[1197] GTGACCATTACCAATAATCGCCTGAATAACAGCAAGATCTATATTAACGGTCGTCTG

[1198] ATTGACCAGAAACCGATTAGTAATCTGGGTAATATTCATGCGAGCAACAACATCAT

[1199] GTTTAAACTGGATGGTTGTCGTGATACCCATCGTTATATTTGGATCAAGTACTTCAA

[1200] CCTGTTCGATAAAGAGTTGAACGAAAAAGAAATTAAAGACCTGTATGATAACCAGA

[1201] GCAACAGCGGTATTCTGAAGGATTTTTGGGGAGATTATCTGCAGTATGACAAACCG

[1202] TATTATATGCTGAATCTGTACGACCCGAATAAATACGTGGATGTGAATAATGTTGGC

[1203] ATCCGTGGTTATATGTACCTGAAAGGTCCGCGTGGTAGCGTTATGACCACAAACATT

[1204] TATCTGAATAGCAGCCTGTATCGCGGAACCAAATTCATCATTAAAAAGTATGCCAG

[1205] CGGCAACAAGGATAATATTGTGCGTAATAATGATCGCGTGTACATTAACGTTGTGG

[1206] TGAAGAATAAAGAATATCGCCTGGCAACCAATGCAAGCCAGGCAGGCGTTGAAAA

[1207] AATTCTGAGTGCCCTGGAAATTCCGGATGTTGGTAATCTGAGCCAGGTTGTTGTGAT

[1208] GAAAAGCAAAAATGATCAGGGCATCACCAACAAGTGCAAAATGAATCTGCAGGAC

[1209] AATAACGGCAACGATATTGGTTTTATTGGCTTCCACCAGTTCAACAATATTGCGAAA

[1210] CTGGTTGCAAGCAATTGGTATAATCGTCAGATTGAACGTAGCAGTCGTACCCTGGGT

[1211] TGTAGCTGGGAATTTATCCCTGTGGATGATGGTTGGGGTGAACGTCCGCTGGAAAA

[1212] CCTGTATTTTCAAGGTGCAAGTCATCATCACCATCACCACCATCATTAA

[1213] The polypeptide sequence (His-tag) of SEQ ID NO:18 -rBoNT / A(0)

[1214] MPFVNKQFNYKDPVNGVDIAYIKIPNAGQMQPVKAFKIHNKIWVIPERDTFTNPEEGDL

[1215] NPPPEAKQVPVSYYDSTYLSTDNEKDNYLKGVTKLFERIYSTDLGRMLLTSIVRGIPFW

[1216] GGSTIDTELKVIDTNCINVIQPDGSYRSEELNLVIIGPSADIIQFECKSFGHEVLNLTRNGY

[1217] GSTQYIRFSPDFTFGFEESLEVDTNPLLGAGKFATDPAVTLAHQLIYAGHRLYGIAINPNR

[1218] VFKVNTNAYYEMSGLEVSFEELRTFGGHDAKFIDSLQENEFRLYYYNKFKDIASTLNKA

[1219] KSIVGTTASLQYMKNVFKEKYLLSEDTSGKFSVDKLKFDKLYKMLTEIYTEDNFVKFFK

[1220] VLNRKTYLNFDKAVFKINIVPKVNYTIYDGFNLRNTNLAANFNGQNTEINNMNFTKLK

[1221] NFTGLFEFYKLLCVRGIITSKTKSLDKGYNKALNDLCIKVNNWDLFFSPSEDNFTNDLN

[1222] KGEEITSDTNIEAAEENISLDLIQQYYLTFNFDNEPENISIENLSSDIIGQLELMPNIERFPN

[1223] GKKYELDKYTMFHYLRAQEFEHGKSRIALTNSVNEALLNPSRVYTFFSSDYVKKVNKA

[1224] TEAAMFLGWVEQLVYDFTDETSEVSTTDKIADITIIIPYIGPALNIGNMLYKDDFVGALIF

[1225] SGAVILLEFIPEIAIPVLGTFALVSYIANKVLTVQTIDNALSKRNEKWDEVYKYIVTNWL

[1226] AKVNTQIDLIRKKMKEALENQAEATKAIINYQYNQYTEEEKNNINFNIDDLSSKLNESIN

[1227] KAMININKFLNQCSVSYLMNSMIPYGVKRLEDFDASLKDALLKYIYDNRGTLIGQVDRL

[1228] KDKVNNTLSTDIPFQLSKYVDNQRLLSTFTEYIKNIINTSILNLRYESNHLIDLSRYASKIN

[1229] IGSKVNFDPIDKNQIQLFNLESSKIEVILKNAIVYNSMYENFSTSFWIRIPKYFNSISLNNE

[1230] YTIINCMENNSGWKVSLNYGEIIWTLQDTQEIKQRVVFKYSQMINISDYINRWIFVTITN

[1231] NRLNNSKIYINGRLIDQKPISNLGNIHASNNIMFKLDGCRDTHRYIWIKYFNLFDKELNE

[1232] KEIKDLYDNQSNSGILKDFWGDYLQYDKPYYMLNLYDPNKYVDVNNVGIRGYMYLK

[1233] GPRGSVMTTNIYLNSSLYRGTKFIIKKYASGNKDNIVRNNDRVYINVVVKNKEYRLATN

[1234] ASQAGVEKILSALEIPDVGNLSQVVVMKSKNDQGITNKCKMNLQDNNGNDIGFIGFHQ

[1235] FNNIAKLVASNWYNRQIERSSRTLGCSWEFIPVDDGWGERPLENLYFQGASHHHHHHH

[1236] H

[1237] SEQ ID NO:19-rLH N The nucleotide sequence of / A (His-tag)

[1238] ATGCCGTTTGTGAACAAGCAGTTCAACTATAAAGATCCGGTTAATGGTGTGGATATC

[1239] GCCTATATCAAAATTCCGAATGCAGGTCAGATGCAGCCGGTTAAAGCCTTTAAAAT

[1240] CCATAACAAAATTTGGGTGATTCCGGAACGTGATACCTTTACCAATCCGGAAGAAG

[1241] GTGATCTGAATCCGCCTCCGGAAGCAAAACAGGTTCCGGTTAGCTATTATGATAGC

[1242] ACCTATCTGAGCACCGATAACGAGAAAGATAACTATCTGAAAGGTGTGACCAAACT

[1243] GTTTGAACGCATTTATAGTACCGATCTGGGTCGTATGCTGCTGACCAGCATTGTTCG

[1244] TGGTATTCCGTTTTGGGGTGGTAGCACCATTGATACCGAACTGAAAGTTATTGACAC

[1245] CAACTGCATTAATGTGATTCAGCCGGATGGTAGCTATCGTAGCGAAGAACTGAATC

[1246] TGGTTATTATTGGTCCGAGCGCAGATATCATTCAGTTTGAATGTAAATCCTTTGGCC

[1247] ACGAAGTTCTGAATCTGACCCGTAATGGTTATGGTAGTACCCAGTATATTCGTTTCA

[1248] GTCCGGATTTTACCTTTGGCTTTGAAGAAAGCCTGGAAGTTGATACAAATCCGCTGT

[1249] TAGGTGCAGGTAAATTTGCAACCGATCCGGCAGTTACCCTGGCACATGAACTGATT

[1250] CATGCCGGTCATCGTCTGTATGGTATTGCAATTAATCCGAACCGTGTGTTCAAAGTG

[1251] AATACCAACGCATATTATGAAATGAGCGGTCTGGAAGTGTCATTTGAAGAACTGCG

[1252] TACCTTTGGTGGTCATGATGCCAAATTTATCGATAGCCTGCAAGAAAATGAATTTCG

[1253] CCTGTACTACTATAACAAATTCAAGGATATTGCGAGCACCCTGAATAAAGCCAAAA

[1254] GCATTGTTGGCACCACCGCAAGCCTGCAGTATATGAAAAATGTGTTTAAAGAAAAA

[1255] TATCTGCTGAGCGAAGATACCAGCGGTAAATTTAGCGTTGACAAACTGAAATTCGA

[1256] TAAACTGTACAAGATGCTGACCGAGATTTATACCGAAGATAACTTCGTGAAGTTTTT

[1257] CAAAGTGCTGAACCGCAAAACCTACCTGAACTTTGATAAAGCCGTGTTCAAAATCA

[1258] ACATCGTGCCGAAAGTGAACTATACCATCTATGATGGTTTTAACCTGCGCAATACCA

[1259] ATCTGGCAGCAAACTTTAATGGTCAGAACACCGAAATCAACAACATGAACTTTACC

[1260] AAACTGAAGAACTTCACCGGTCTGTTCGAATTTTACAAACTGCTGTGTGTTCGTGGC

[1261] ATTATTACCAGCAAAACCAAAAGTCTGGATAAAGGCTACAATAAAGCCCTGAATGA

[1262] TCTGTGCATTAAGGTGAATAATTGGGACCTGTTTTTTAGCCCGAGCGAGGATAATTT

[1263] CACCAACGATCTGAACAAAGGCGAAGAAATTACCAGCGATACCAATATTGAAGCA

[1264] GCCGAAGAAAACATTAGCCTGGATCTGATTCAGCAGTATTATCTGACCTTCAACTTC

[1265] GATAATGAGCCGGAAAATATCAGCATTGAAAACCTGAGCAGCGATATTATTGGCCA

[1266] GCTGGAACTGATGCCGAATATTGAACGTTTTCCGAACGGCAAAAAATACGAGCTGG

[1267] ATAAATACACCATGTTCCATTATCTGCGTGCCCAAGAATTTGAACATGGTAAAAGCC

[1268] GTATTGCACTGACCAATAGCGTTAATGAAGCACTGCTGAACCCGAGCCGTGTTTATA

[1269] CCTTTTTTAGCAGCGATTACGTGAAAAAGGTTAACAAAGCAACCGAAGCAGCCATG

[1270] TTTTTAGGTTGGGTTGAACAGCTGGTTTATGATTTCACCGATGAAACCAGCGAAGTT

[1271] AGCACCACCGATAAAATTGCAGATATTACCATCATCATCCCGTATATCGGTCCGGCA

[1272] CTGAATATTGGCAATATGCTGTATAAAGACGATTTTGTGGGTGCCCTGATTTTTAGC

[1273] GGTGCAGTTATTCTGCTGGAATTTATTCCGGAAATTGCCATTCCGGTTCTGGGCACC

[1274] TTTGCACTGGTGAGCTATATTGCAAATAAAGTTCTGACCGTGCAGACCATCGATAAT

[1275] GCACTGAGCAAACGTAACGAAAAATGGGATGAAGTGTACAAGTATATCGTGACCA

[1276] ATTGGCTGGCAAAAGTTAACACCCAGATTGACCTGATTCGCAAGAAGATGAAAGAA

[1277] GCACTGGAAAATCAGGCAGAAGCAACCAAAGCCATTATCAACTATCAGTATAACCA

[1278] GTACACCGAAGAAGAGAAAAATAACATCAACTTCAACATCGACGATCTGTCCAGCA

[1279] AACTGAACGAAAGCATCAACAAAGCCATGATTAACATTAACAAATTTCTGAACCAG

[1280] TGCAGCGTGAGCTATCTGATGAATAGCATGATTCCGTATGGTGTGAAACGTCTGGA

[1281] AGATTTTGATGCAAGCCTGAAAGATGCCCTGCTGAAATATATCTATGATAATCGTGG

[1282] CACCCTGATTGGTCAGGTTGATCGTCTGAAAGATAAAGTGAACAACACCCTGAGTA

[1283] CCGATATTCCTTTTCAGCTGAGCAAATATGTGGATAATCAGCGTCTGCTGTCAACCG

[1284] AAAATCTGTATTTCCAGGGTGCAAGTCATCATCACCATCACCACCATCATTAA

[1285] SEQ ID NO:20-rLH N / A polypeptide sequence (His-tag)

[1286] MPFVNKQFNYKDPVNGVDIAYIKIPNAGQMQPVKAFKIHNKIWVIPERDTFTNPEEGDL

[1287] NPPPEAKQVPVSYYDSTYLSTDNEKDNYLKGVTKLFERIYSTDLGRMLLTSIVRGIPFW

[1288] GGSTIDTELKVIDTNCINVIQPDGSYRSEELNLVIIGPSADIIQFECKSFGHEVLNLTRNGY

[1289] GSTQYIRFSPDFTFGFEESLEVDTNPLLGAGKFATDPAVTLAHELIHAGHRLYGIAINPNR

[1290] VFKVNTNAYYEMSGLEVSFEELRTFGGHDAKFIDSLQENEFRLYYYNKFKDIASTLNKA

[1291] KSIVGTTASLQYMKNVFKEKYLLSEDTSGKFSVDKLKFDKLYKMLTEIYTEDNFVKFFK

[1292] VLNRKTYLNFDKAVFKINIVPKVNYTIYDGFNLRNTNLAANFNGQNTEINNMNFTKLK

[1293] NFTGLFEFYKLLCVRGIITSKTKSLDKGYNKALNDLCIKVNNWDLFFSPSEDNFTNDLN

[1294] KGEEITSDTNIEAAEENISLDLIQQYYLTFNFDNEPENISIENLSSDIIGQLELMPNIERFPN

[1295] GKKYELDKYTMFHYLRAQEFEHGKSRIALTNSVNEALLNPSRVYTFFSSDYVKKVNKA

[1296] TEAAMFLGWVEQLVYDFTDETSEVSTTDKIADITIIIPYIGPALNIGNMLYKDDFVGALIF

[1297] SGAVILLEFIPEIAIPVLGTFALVSYIANKVLTVQTIDNALSKRNEKWDEVYKYIVTNWL

[1298] AKVNTQIDLIRKKMKEALENQAEATKAIINYQYNQYTEEEKNNINFNIDDLSSKLNESIN

[1299] KAMININKFLNQCSVSYLMNSMIPYGVKRLEDFDASLKDALLKYIYDNRGTLIGQVDRL

[1300] KDKVNNTLSTDIPFQLSKYVDNQRLLSTENLYFQGASHHHHHHHH

[1301] SEQ ID NO:21-rH C The nucleotide sequence of / A (His-tag)

[1302] ATGCATCATCACCATCACCACGAAAATCTATACTTCCAAGGAAAAAACATCATCAA

[1303] TACTAGCATTCTGAACCTGCGTTACGAGAGCAATCATCTGATTGATCTGAGCCGTTA

[1304] TGCAAGCAAGATCAACATCGGTAGCAAGGTCAATTTTGACCCGATCGATAAGAACC

[1305] AGATCCAGCTGTTTAATCTGGAATCGAGCAAAATTGAGGTTATCCTGAAAAACGCC

[1306] ATTGTCTACAACTCCATGTACGAGAATTTCTCCACCAGCTTCTGGATTCGCATCCCG

[1307] AAATACTTCAACAGCATTAGCCTGAACAACGAGTATACTATCATCAACTGTATGGA

[1308] GAACAACAGCGGTTGGAAGGTGTCTCTGAACTATGGTGAGATCATTTGGACCTTGC

[1309] AGGACACCCAAGAGATCAAGCAGCGCGTCGTGTTCAAGTACTCTCAAATGATCAAC

[1310] ATTTCCGATTACATTAATCGTTGGATCTTCGTGACCATTACGAATAACCGTCTGAAT

[1311] AACAGCAAGATTTACATCAATGGTCGCTTGATCGATCAGAAACCGATTAGCAACCT

[1312] GGGTAATATCCACGCAAGCAACAACATTATGTTCAAATTGGACGGTTGCCGCGATA

[1313] CCCATCGTTATATCTGGATCAAGTATTTCAACCTGTTTGATAAAGAACTGAATGAGA

[1314] AGGAGATCAAAGATTTGTATGACAACCAATCTAACAGCGGCATTTTGAAGGACTTC

[1315] TGGGGCGATTATCTGCAATACGATAAGCCGTACTATATGCTGAACCTGTATGATCCG

[1316] AACAAATATGTGGATGTCAATAATGTGGGTATTCGTGGTTACATGTATTTGAAGGGT

[1317] CCGCGTGGCAGCGTTATGACGACCAACATTTACCTGAACTCTAGCCTGTACCGTGGT

[1318] ACGAAATTCATCATTAAGAAATATGCCAGCGCAACAAAGATAACATTGTGCGTAA

[1319] TAACGATCGTGTCTACATCAACGTGGTCGTGAAGAATAAAGAGTACCGTCTGGCGA

[1320] CCAACGCTTCGCAGGCGGTGTTGAGAAAATTCTGAGCGCGTTGGAGATCCCTGAT

[1321] GTCGGTAATCTGAGCCAAGTCGTGGTTATGAAGAGCAAGAACGACCAGGGTATCAC

[1322] TAACAAGTGCAAGATGAACCTGCAAGACAACAATGGTAACGACATCGGCTTATTG

[1323] GTTTCCACCAGTTCAACAATATTGCTAAACTGGTAGCGAGCAATTGGTACAATCGTC

[1324] AGATTGAGCGCAGCAGCCGTACTTTGGGCTGTAGCTGGGAGTTTATCCCGGTCGAT

[1325] GATGGTTGGGGCGAACGTCCGCTGTAA

[1326] SEQ ID NO:22-rH C / A polypeptide sequence (His-tag)

[1327] MHHHHHHENLYFQGKNIINTSILNLRYESNHLIDLSRYASKINIGSKVNFDPIDKNQIQLF

[1328] NLESSKIEVILKNAIVYNSMYENFSTSFWIRIPKYFNSISLNNEYTIINCMENNSGWKVSL

[1329] NYGEIIWTLQDTQEIKQRVVFKYSQMINISDYINRWIFVTITNNRLNNSKIYINGRLIDQK

[1330] PISNLGNIHASNNIMFKLDGCRDTHRYIWIKYFNLFDKELNEKEIKDLYDNQSNSGILKD

[1331] FWGDYLQYDKPYYMLNLYDPNKYVDVNNVGIRGYMYLKGPRGSVMTTNIYLNSSLY

[1332] RGTKFIIKKYASGNKDNIVRNNDRVYINVVVKNKEYRLATNASQAGVEKILSALEIPDV

[1333] GNLSQVVVMKSKNDQGITNKCKMNLQDNNGNDIGFIGFHQFNNIAKLVASNWYNRQI

[1334] ERSSRTLGCSWEFIPVDDGWGERPL

[1335] Nucleotide sequence (His-tag) of SEQ ID NO:23-rLC / A

[1336] ATGCCGTTTGTGAACAAGCAGTTCAACTATAAAGATCCGGTTAATGGTGTGGATATC

[1337] GCCTATATCAAAATTCCGAATGCAGGTCAGATGCAGCCGGTTAAAGCCTTTAAAAT

[1338] CCATAACAAAATTTGGGTGATTCCGGAACGTGATACCTTTACCAATCCGGAAGAAG

[1339] GTGATCTGAATCCGCCTCCGGAAGCAAAACAGGTTCCGGTTAGCTATTATGATAGC

[1340] ACCTATCTGAGCACCGATAACGAGAAAGATAACTATCTGAAAGGTGTGACCAAACT

[1341] GTTTGAACGCATTTATAGTACCGATCTGGGTCGTATGCTGCTGACCAGCATTGTTCG

[1342] TGGTATTCCGTTTTGGGGTGGTAGCACCATTGATACCGAACTGAAAGTTATTGACAC

[1343] CAACTGCATTAATGTGATTCAGCCGGATGGTAGCTATCGTAGCGAAGAACTGAATC

[1344] TGGTTATTATTGGTCCGAGCGCAGATATCATTCAGTTTGAATGTAAATCCTTTGGCC

[1345] ACGAAGTTCTGAATCTGACCCGTAATGGTTATGGTAGTACCCAGTATATTCGTTTCA

[1346] GTCCGGATTTTACCTTTGGCTTTGAAGAAAGCCTGGAAGTTGATACAAATCCGCTGT

[1347] TAGGTGCAGGTAAATTTGCAACCGATCCGGCAGTTACCCTGGCACATGAACTGATT

[1348] CATGCCGGTCATCGTCTGTATGGTATTGCAATTAATCCGAACCGTGTGTTCAAAGTG

[1349] AATACCAACGCATATTATGAAATGAGCGGTCTGGAAGTGTCATTTGAAGAACTGCG

[1350] TACCTTTGGTGGTCATGATGCCAAATTTATCGATAGCCTGCAAGAAAATGAATTTCG

[1351] CCTGTACTACTATAACAAATTCAAGGATATTGCGAGCACCCTGAATAAAGCCAAAA

[1352] GCATTGTTGGCACCACCGCAAGCCTGCAGTATATGAAAAATGTGTTTAAAGAAAAA

[1353] TATCTGCTGAGCGAAGATACCAGCGGTAAATTTAGCGTTGACAAACTGAAATTCGA

[1354] TAAACTGTACAAGATGCTGACCGAGATTTATACCGAAGATAACTTCGTGAAGTTTTT

[1355] CAAAGTGCTGAACCGCAAAACCTACCTGAACTTTGATAAAGCCGTGTTCAAAATCA

[1356] ACATCGTGCCGAAAGTGAACTATACCATCTATGATGGTTTTAACCTGCGCAATACCA

[1357] ATCTGGCAGCAAACTTTAATGGTCAGAACACCGAAATCAACAACATGAACTTTACC

[1358] AAACTGAAGAACTTCACCGGTCTGTTTGAAGAGAATCTGTATTTCCAGGGTGCAAG

[1359] TCATCATCACCATCACCACCATCATTAA

[1360] The polypeptide sequence (His-tag) of SEQ ID NO:24-rLC / A

[1361] MPFVNKQFNYKDPVNGVDIAYIKIPNAGQMQPVKAFKIHNKIWVIPERDTFTNPEEGDL

[1362] NPPPEAKQVPVSYYDSTYLSTDNEKDNYLKGVTKLFERIYSTDLGRMLLTSIVRGIPFW

[1363] GGSTIDTELKVIDTNCINVIQPDGSYRSEELNLVIIGPSADIIQFECKSFGHEVLNLTRNGY

[1364] GSTQYIRFSPDFTFGFEESLEVDTNPLLGAGKFATDPAVTLAHELIHAGHRLYGIAINPNR

[1365] VFKVNTNAYYEMSGLEVSFEELRTFGGHDAKFIDSLQENEFRLYYYNKFKDIASTLNKA

[1366] KSIVGTTASLQYMKNVFKEKYLLSEDTSGKFSVDKLKFDKLYKMLTEIYTEDNFVKFFK

[1367] VLNRKTYLNFDKAVFKINIVPKVNYTIYDGFNLRNTNLAANFNGQNTEINNMNFTKLK

[1368] NFTGLFEENLYFQGASHHHHHHHH

[1369] Nucleotide sequence (His-tag) of SEQ ID NO:25-rBoNT / FA(0)

[1370] ATGCCGGTTGTGATTAACAGCTTCAATTATGATGATCCGGTGAACGATAACACCATC

[1371] ATTTATATCCGTCCGCCTTATTATGAAACCAGCAACACCTATTTCAAAGCCTTCCAG

[1372] ATTATGGATAACGTGTGGATTATTCCGGAACGTTATCGTCTGGGTATTGATCCGAGC

[1373] CTGTTTAATCCGCCTGTTAGCCTGAAAGCAGGTAGTGATGGTTATTTTGATCCGAAT

[1374] TATCTGAGCACCAACACCGAGAAAAACAAATACCTGCAGATTATGATCAAGCTGTT

[1375] CAAACGCATTAATAGCAAACCGGCAGGTCAGATTCTGCTGGAAGAAATCAAAAATG

[1376] CAATTCCGTATCTGGGCAACAGCTATACCCAAGAAGAACAGTTTACCACCAATAAT

[1377] CGTACCGTGAGCTTTAATGTTAAACTGGCCAATGGTAATATCGTTCAGCAGATGGCA

[1378] AATCTGATTATTTGGGGTCCGGGTCCTGATCTGACCACAAATAAAACCGGTGGTATC

[1379] ATCTATAGCCCGTATCAGAGCATGGAAGCAACCCCGTATAAAGATGGTTTTGGTAG

[1380] CATTATGACCGTGGAATTTAGTCCGGAATATGCAACCGCCTTTAACGATATTTCAAT

[1381] TGCAAGCCATAGTCCGTCGCTGTTTATCAAAGATCCGGCACTGATTCTGATGCACCA

[1382] GCTGATTTATGTTCTGCATGGTCTGTATGGCACCTATATCACCGAATACAAAATTAC

[1383] CCCGAATGTGGTTCAGAGCTATATGAAAGTTACCAAACCGATTACCAGCGCAGAAT

[1384] TTCTGACCTTTGGTGGTCGTGATCGCAATATTGTTCCGCAGAGCATTCAGAGCCAGC

[1385] TGTATAACAAAGTTCTGAGCGATTATAAACGTATTGCCAGCCGTCTGAATAAAGTTA

[1386] ATACCGCAACCGCACTGATCAACATCGATGAATTCAAAAACCTGTACGAGTGGAAA

[1387] TACCAGTTTGCCAAAGATAGCAATGGTGTGTATAGCGTGGATCTGAACAAATTTGA

[1388] GCAGCTGTACAAAAAAATCTATAGCTTCACCGAATTCAACCTGGCCTATGAGTTTAA

[1389] AATCAAAACCCGTCTGGGTTATCTGGCCGAAAATTTTGGTCCGTTTTATCTGCCGAA

[1390] TCTGCTGGATGATAGCATTTATACCGAAGTGGATGGTTTTAACATTGGTGCACTGAG

[1391] CATTAACTATCAGGGTCAGAATATTGGCAGCGATATCAACAGCATCAAAAAACTGC

[1392] AAGGTCAGGGTGTTGTTAGCCGTGTTGTTCGTCTGTGTAGCAATAGCAATACCAAAA

[1393] ACAGCCTGTGCATTACCGTTAATAATCGCGACCTGTTTTTTATCGCAAGCCAAGAAA

[1394] GCTATGGCGAGAATACCATTAACACCTATAAAGAGATTGACGATACCACCACACTG

[1395] GATCCGAGCTTTGAAGATATTCTGGATAAAGTGATCCTGAACTTCAACGAACAGGT

[1396] TATTCCGCAGATGCCGAATCGTAATGTTAGCACCGATATTCAGAAAGACAACTACA

[1397] TCCCGAAATACGATTATAACCGCACCGACATTATCGATAGCTATGAAGTTGGTCGC

[1398] AACTACAACACCTTTTTCTATCTGAATGCCCAGAAATTTAGCCCGAACGAAAGCAAT

[1399] ATTACCCTGACCAGCAGCTTTGATACAGGTCTGTTAGAAGGTAGCAAAGTGTATAC

[1400] CTTTTTCAGCAGCGATTTCATTAACAACATCAACAAACCGGTTCAGGCCCTGCTGTT

[1401] TATTGAATGGGTTAAACAGGTGATTCGCGATTTTACCACCGAAGCAACCAAAACCT

[1402] CAACCGTTGATAAACTGAAAGATATTAGCCTGGTGGTGCCGTATATTGGTCTGGCAC

[1403] TGAATATTGGTGATGAGATCTACAAACAGCATTTTGCAGAAGCAGTTGAACTGGTT

[1404] GGTGCAGGTCTGCTGCTGGAATTTTCACCGGAATTTCTTATTCCGACGCTGCTGATT

[1405] TTTACCATCAAAGGTTATCTGACCGGTAGCATTCGCGATAAAGACAAAATCATTAA

[1406] AACCCTGGATAACGCCCTGAATGTTCGTGATCAGAAATGGAAAGAACTGTATCGTT

[1407] GGGTTGTTAGCAAATGGCTGACCACCATTAATACGCAGTTCAACAAACGCAAAGAA

[1408] CAAATGTACAAAGCCCTGAAAAATCAGGCCACCGCCATTAAAAAGATCATCGAGAA

[1409] CAAATATAACAACTATACCACCGATGAAAAAAGCAAGATCGATAGCAGCTATAACA

[1410] TCAACGAAATTGAACGCACCCTGAACGAAAAAATCAATCTGGCCATGAAAAACATC

[1411] GAGCAGTTTATTACCGAAAGCAGCATTGCCTATCTGATCAATATCATCAACAACGA

[1412] AACGATCCAGAAACTGAAAAGCTATGATGACCTGGTTCGTCGTTATCTGCTGGGTTA

[1413] TATTCGTAATCATAGCAGCATTCTGGGCAATAGCGTTGAAGAACTGAATTCCAAAG

[1414] TGAACAACCATCTGGATAATGGCATTCCGTTTGAACTGAGCAGTTATACCAATGATA

[1415] GCCTGCTGATCCGCTACTTCAATAAAAACTATGGCGAACTGAAGTACAACTGCATTC

[1416] TGAACATCAAATATGAGATGGATCGTGACAAACTGGTTGATAGCAGCGGTTATCGT

[1417] AGCCGTATCAATATTGGTACAGGCGTCAAATTTAGCGAGATCGATAAAAATCAAGT

[1418] GCAGCTGAGCAATCTGGAATCCAGCAAAATTGAAGTCATTCTGAATAACGGCGTCA

[1419] TCTATAACAGCATGTATGAAAACTTTTCGACCAGCTTTTGGATTCGCATTCCGAAAT

[1420] ACTTTCGCAACATCAATAACGAGTACAAGATCATCAGCTGTATGCAGAATAATAGC

[1421] GGTTGGGAAGTGAGCCTGAATTTTAGCAATATGAACTCGAAAATCATCTGGACCCT

[1422] GCAGGATACCGAAGGTATCAAAAAAACCGTTGTGTTTCAGTACACCCAGAACATTA

[1423] ACATTAGCGACTATATCAACCGCTGGATCTTTGTGACCATTACAAATAATCGTCTGA

[1424] GCAACAGCAAAATCTACATTAATGGTCGCCTGATCAACGAAGAAAGCATTAGCGAT

[1425] CTGGGTAATATCCATGCCAGCAACAACATTATGTTTAAACTGGATGGTTGCCGTGAT

[1426] CCGCATCGTTATATCTGGATTAAATACTTTAACCTGTTTGACAAAGAGCTGAACAAG

[1427] AAAGAAATTAAAGATCTGTACGACAACCAGAGCAATAGCGGTATTCTGAAAGATTT

[1428] CTGGGGTGATTATCTGCAGTATGACAAACCGTATTATATGCTGAATCTGTATGACCC

[1429] GAATAAGTATCTGGATGTGAATAATGTTGGCATCCGTGGCTATATGTATCTGAAAG

[1430] GTCCGCGTGGTCGTATTGTGACCACCAACATTTATCTGAATAGCACCCTGTATATGG

[1431] GCACCAAATTCATCATTAAGAAATATGCCAGCGGCAACAAAGATAACATTGTGCGT

[1432] AATAATGATCGCGTGTATATTAACGTGGTGGTGAAGAATAAAGAATATCGCCTGGC

[1433] AACCAATGCAAGCCAGGCAGGCGTTGAAAAAATTCTGAGCGCAGTTGAAATCCCGG

[1434] ATGTTGGTAATCTGAGCCAGGTTGTTGTGATGAAAAGCGAAAATGATCAGGGCATT

[1435] CGCAACAAGTGTAAAATGAATCTGCAAGACAATAACGGCAACGATATTGGCTTTAT

[1436] CGGCTTTCACCAGTTTAATAACATTGCAAAACTGGTGGCCAGCAACTGGTATAACC

[1437] GTCAGATTGGTAAAGCAAGCCGTACCTTTGGTTGTAGCTGGGAATTTATCCCGGTTG

[1438] ATGATGGTTGGGGTGAAAGCAGCCTGGAAAATCTGTATTTCCAGGGTGCCAGTCAT

[1439] CATCACCACCATCACCATCACTGA

[1440] The polypeptide sequence (His-tag) of SEQ ID NO:26-rBoNT / FA(0)

[1441] MPVVINSFNYDDPVNDNTIIYIRPPYYETSNTYFKAFQIMDNVWIIPERYRLGIDPSLFNPP

[1442] VSLKAGSDGYFDPNYLSTNTEKNKYLQIMIKLFKRINSKPAGQILLEEIKNAIPYLGNSYT

[1443] QEEQFTTNNRTVSFNVKLANGNIVQQMANLIIWGPGPDLTTNKTGGIIYSPYQSMEATP

[1444] YKDGFGSIMTVEFSPEYATAFNDISIASHSPSLFIKDPALILMHQLIYVLHGLYGTYITEYK

[1445] ITPNVVQSYMKVTKPITSAEFLTFGGRDRNIVPQSIQSQLYNKVLSDYKRIASRLNKVNT

[1446] ATALINIDEFKNLYEWKYQFAKDSNGVYSVDLNKFEQLYKKIYSFTEFNLAYEFKIKTR

[1447] LGYLAENFGPFYLPNLLDDSIYTEVDGFNIGALSINYQGQNIGSDINSIKKLQGQGVVSR

[1448] VVRLCSNSNTKNSLCITVNNRDLFFIASQESYGENTINTYKEIDDTTTLDPSFEDILDKVIL

[1449] NFNEQVIPQMPNRNVSTDIQKDNYIPKYDYNRTDIIDSYEVGRNYNTFFYLNAQKFSPNE

[1450] SNITLTSSFDTGLLEGSKVYTFFSSDFINNINKPVQALLFIEWVKQVIRDFTTEATKTSTVD

[1451] KLKDISLVVPYIGLALNIGDEIYKQHFAEAVELVGAGLLLEFSPEFLIPTLLIFTIKGYLTG

[1452] SIRDKDKIIKTLDNALNVRDQKWKELYRWVVSKWLTTINTQFNKRKEQMYKALKNQA

[1453] TAIKKIIENKYNNYTTDEKSKIDSSYNINEIERTLNEKINLAMKNIEQFITESSIAYLINIINN

[1454] ETIQKLKSYDDLVRRYLLGYIRNHSSILGNSVEELNSKVNNHLDNGIPFELSSYTNDSLLI

[1455] RYFNKNYGELKYNCILNIKYEMDRDKLVDSSGYRSRINIGTGVKFSEIDKNQVQLSNLE

[1456] SSKIEVILNNGVIYNSMYENFSTSFWIRIPKYFRNINNEYKIISCMQNNSGWEVSLNFSNM

[1457] NSKIIWTLQDTEGIKKTVVFQYTQNINISDYINRWIFVTITNNRLSNSKIYINGRLINEESIS

[1458] DLGNIHASNNIMFKLDGCRDPHRYIWIKYFNLFDKELNKKEIKDLYDNQSNSGILKDFW

[1459] GDYLQYDKPYYMLNLYDPNKYLDVNNVGIRGYMYLKGPRGRIVTTNIYLNSTLYMGT

[1460] KFIIKKYASGNKDNIVRNNDRVYINVVVKNKEYRLATNASQAGVEKILSAVEIPDVGNL

[1461] SQVVVMKSENDQGIRNKCKMNLQDNNGNDIGFIGFHQFNNIAKLVASNWYNRQIGKA

[1462] SRTFGCSWEFIPVDDGWGESSLENLYFQGASHHHHHHHH

[1463] SEQ ID NO:27-rLH N / FA nucleotide sequence (His-tag)

[1464] ATGCCGGTTGTGATTAACAGCTTCAATTATGATGATCCGGTGAACGATAACACCATC

[1465] ATTTATATCCGTCCGCCTTATTATGAAACCAGCAACACCTATTTCAAAGCCTTCCAG

[1466] ATTATGGATAACGTGTGGATTATTCCGGAACGTTATCGTCTGGGTATTGATCCGAGC

[1467] CTGTTTAATCCGCCTGTTAGCCTGAAAGCAGGTAGTGATGGTTATTTTGATCCGAAT

[1468] TATCTGAGCACCAACACCGAGAAAAACAAATACCTGCAGATTATGATCAAGCTGTT

[1469] CAAACGCATTAATAGCAAACCGGCAGGTCAGATTCTGCTGGAAGAAATCAAAAATG

[1470] CAATTCCGTATCTGGGCAACAGCTATACCCAAGAAGAACAGTTTACCACCAATAAT

[1471] CGTACCGTGAGCTTTAATGTTAAACTGGCCAATGGTAATATCGTTCAGCAGATGGCA

[1472] AATCTGATTATTTGGGGTCCGGGTCCTGATCTGACCACAAATAAAACCGGTGGTATC

[1473] ATCTATAGCCCGTATCAGAGCATGGAAGCAACCCCGTATAAAGATGGTTTTGGTAG

[1474] CATTATGACCGTGGAATTTAGTCCGGAATATGCAACCGCCTTTAACGATATTTCAAT

[1475] TGCAAGCCATAGTCCGTCGCTGTTTATCAAAGATCCGGCACTGATTCTGATGCATGA

[1476] ACTGATTCATGTTCTGCATGGTCTGTATGGCACCTATATTACCGAATACAAAATTAC

[1477] CCCGAATGTGGTGCAGAGCTATATGAAAGTTACCAAACCGATTACCAGCGCAGAAT

[1478] TTCTGACCTTTGGTGGTCGTGATCGCAATATTGTTCCGCAGAGCATTCAGAGCCAGC

[1479] TGTATAACAAAGTTCTGAGCGATTATAAACGTATTGCCAGCCGTCTGAATAAAGTTA

[1480] ATACCGCAACCGCACTGATCAACATCGATGAATTCAAAAACCTGTACGAGTGGAAA

[1481] TACCAGTTTGCCAAAGATAGCAATGGTGTGTATAGCGTGGATCTGAACAAATTTGA

[1482] GCAGCTGTACAAAAAAATCTATAGCTTCACCGAATTCAACCTGGCCTATGAGTTTAA

[1483] AATCAAAACCCGTCTGGGTTATCTGGCCGAAAATTTTGGTCCGTTTTATCTGCCGAA

[1484] TCTGCTGGATGATAGCATTTATACCGAAGTGGATGGTTTTAACATTGGTGCACTGAG

[1485] CATTAACTATCAGGGTCAGAATATTGGCAGCGATATCAACAGCATCAAAAAACTGC

[1486] AAGGTCAGGGTGTTGTTAGCCGTGTTGTTCGTCTGTGTAGCAATAGCAATACCAAAA

[1487] ACAGCCTGTGCATTACCGTTAATAATCGCGACCTGTTTTTTATCGCAAGCCAAGAAA

[1488] GCTATGGCGAGAATACCATTAACACCTATAAAGAGATTGACGATACCACCACACTG

[1489] GATCCGAGCTTTGAAGATATTCTGGATAAAGTGATCCTGAACTTCAACGAACAGGT

[1490] TATTCCGCAGATGCCGAATCGTAATGTTAGCACCGATATTCAGAAAGACAACTACA

[1491] TCCCGAAATACGATTATAACCGCACCGACATTATCGATAGCTATGAAGTTGGTCGC

[1492] AACTACAACACCTTTTTCTATCTGAATGCCCAGAAATTTAGCCCGAACGAAAGCAAT

[1493] ATTACCCTGACCAGCAGCTTTGATACAGGTCTGTTAGAAGGTAGCAAAGTGTATAC

[1494] CTTTTTCAGCAGCGATTTCATTAACAACATCAACAAACCGGTTCAGGCCCTGCTGTT

[1495] TATTGAATGGGTTAAACAGGTGATTCGCGATTTTACCACCGAAGCAACCAAAACCT

[1496] CAACCGTTGATAAACTGAAAGATATTAGCCTGGTGGTGCCGTATATTGGTCTGGCAC

[1497] TGAATATTGGTGATGAGATCTACAAACAGCATTTTGCAGAAGCAGTTGAACTGGTT

[1498] GGTGCAGGTCTGCTGCTGGAATTTTCACCGGAATTTCTTATTCCGACGCTGCTGATT

[1499] TTTACCATCAAAGGTTATCTGACCGGTAGCATTCGCGATAAAGACAAAATCATTAA

[1500] AACCCTGGATAACGCCCTGAATGTTCGTGATCAGAAATGGAAAGAACTGTATCGTT

[1501] GGGTTGTTAGCAAATGGCTGACCACCATTAATACGCAGTTCAACAAACGCAAAGAA

[1502] CAAATGTACAAAGCCCTGAAAAATCAGGCCACCGCCATTAAAAAGATCATCGAGAA

[1503] CAAATATAACAACTATACCACCGATGAAAAAAGCAAGATCGATAGCAGCTATAACA

[1504] TCAACGAAATTGAACGCACCCTGAACGAAAAAATCAATCTGGCCATGAAAAACATC

[1505] GAGCAGTTTATTACAGAAAGCAGCATTGCCTACCTGATCAATATCATCAACAACGA

[1506] AACCATTCAGAAACTGAAAAGCTATGATGACCTGGTTCGTCGTTATCTGCTGGGTTA

[1507] TATTCGTAATCATAGCAGCATTCTGGGCAATAGCGTTGAAGAACTGAATTCCAAAG

[1508] TGAACAACCATCTGGATAATGGCATTCCGTTTGAACTGAGCAGTTATACCAATGATA

[1509] GCCTGCTGATCCGCTACTTCAATAAAAACTATGGCGAAGAGAACCTGTATTTCCAG

[1510] GGTGCCAGTCATCATCACCACCATCACCATCACTGA

[1511] SEQ ID NO:28-rLH N / FA polypeptide sequence (His-tag)

[1512] MPVVINSFNYDDPVNDNTIIYIRPPYYETSNTYFKAFQIMDNVWIIPERYRLGIDPSLFNPP

[1513] VSLKAGSDGYFDPNYLSTNTEKNKYLQIMIKLFKRINSKPAGQILLEEIKNAIPYLGNSYT

[1514] QEEQFTTNNRTVSFNVKLANGNIVQQMANLIIWGPGPDLTTNKTGGIIYSPYQSMEATP

[1515] YKDGFGSIMTVEFSPEYATAFNDISIASHSPSLFIKDPALILMHELIHVLHGLYGTYITEYK

[1516] ITPNVVQSYMKVTKPITSAEFLTFGGRDRNIVPQSIQSQLYNKVLSDYKRIASRLNKVNT

[1517] ATALINIDEFKNLYEWKYQFAKDSNGVYSVDLNKFEQLYKKIYSFTEFNLAYEFKIKTR

[1518] LGYLAENFGPFYLPNLLDDSIYTEVDGFNIGALSINYQGQNIGSDINSIKKLQGQGVVSR

[1519] VVRLCSNSNTKNSLCITVNNRDLFFIASQESYGENTINTYKEIDDTTTLDPSFEDILDKVIL

[1520] NFNEQVIPQMPNRNVSTDIQKDNYIPKYDYNRTDIIDSYEVGRNYNTFFYLNAQKFSPNE

[1521] SNITLTSSFDTGLLEGSKVYTFFSSDFINNINKPVQALLFIEWVKQVIRDFTTEATKTSTVD

[1522] KLKDISLVVPYIGLALNIGDEIYKQHFAEAVELVGAGLLLEFSPEFLIPTLLIFTIKGYLTG

[1523] SIRDKDKIIKTLDNALNVRDQKWKELYRWVVSKWLTTINTQFNKRKEQMYKALKNQA

[1524] TAIKKIIENKYNNYTTDEKSKIDSSYNINEIERTLNEKINLAMKNIEQFITESSIAYLINIINN

[1525] ETIQKLKSYDDLVRRYLLGYIRNHSSILGNSVEELNSKVNNHLDNGIPFELSSYTNDSLLI

[1526] RYFNKNYGEENLYFQGASHHHHHHHH

[1527] SEQ ID NO:29-rH C / FA nucleotide sequence (His-tag)

[1528] ATGCTGAAGTATAACTGCATCCTGAACATCAAATATGAGATGGATCGTGATAAACT

[1529] GGTTGATAGCAGCGGTTATCGTAGCCGTATCAATATTGGCACCGGTGTGAAATTTAG

[1530] CGAGATCGATAAAAATCAGGTGCAGCTGAGCAATCTGGAAAGCAGCAAAATTGAA

[1531] GTGATTCTGAATAACGGCGTGATCTACAATAGCATGTATGAAAACTTTTCGACCAGC

[1532] TTCTGGATTCGCATTCCGAAATACTTTCGCAACATCAACAACGAGTACAAGATTATC

[1533] AGCTGTATGCAGAATAATAGCGGTTGGGAAGTTAGCCTGAATTTCAGCAATATGAA

[1534] CAGCAAAATCATTTGGACCCTGCAGGATACCGAAGGTATCAAAAAAACCGTTGTGT

[1535] TTCAGTACACCCAGAACATTAACATCAGCGATTACATTAACCGCTGGATCTTTGTGA

[1536] CCATTACCAATAATCGTCTGAGCAACAGCAAGATCTATATTAACGGTCGCCTGATTA

[1537] ACGAAGAGAGCATTAGCGATCTGGGTAATATTCATGCCAGCAACAACATCATGTTT

[1538] AAACTGGATGGTTGTCGTGATCCGCATCGTTATATTTGGATCAAATACTTCAACCTG

[1539] TTTGATAAAGAACTGAACAAAAAAGAAATCAAAGACCTGTATGATAACCAGAGCA

[1540] ATAGCGGCATTCTGAAAGATTTTTGGGGTGATTATCTGCAGTATGACAAACCGTATT

[1541] ACATGCTGAATCTGTACGATCCGAACAAATATCTGGATGTGAATAATGTGGGTATC

[1542] CGTGGCTATATGTATCTGAAAGGTCCGCGTGGTCGTATTGTTACCACCAACATTTAT

[1543] CTGAATAGCACCCTGTATATGGGCACCAAATTCATCATTAAAAAGTATGCCAGCGG

[1544] CAACAAAGATAACATTGTGCGTAATAATGATCGCGTGTATATCAATGTGGTGGTGA

[1545] AGAATAAAGAATATCGTCTGGCCACCAATGCAAGCCAGGCAGGCGTTGAAAAAATT

[1546] CTGAGCGCAGTTGAAATTCCGGATGTTGGTAATCTGAGCCAGGTTGTTGTTATGAAA

[1547] AGCGAAAATGATCAGGGCATTCGCAACAAATGCAAAATGAATCTGCAGGACAATA

[1548] ACGGCAACGATATTGGTTTTATTGGCTTCCACCAGTTCAACAACATTGCAAAACTGG

[1549] TGGCGAGCAATTGGTATAATCGTCAGATTGGTAAAGCAAGCCGTACCTTTGGTTGTA

[1550] GCTGGGAATTTATTCCGGTTGATGATGGTTGGGGTGAAAGCAGCCTGGAAAATCTG

[1551] TATTTTCAGGGTGCAAGTCATCATCACCACCATCACCATCATTAA

[1552] SEQ ID NO:30-rH C / FA polypeptide sequence (His-tag)

[1553] MLKYNCILNIKYEMDRDKLVDSSGYRSRINIGTGVKFSEIDKNQVQLSNLESSKIEVILN

[1554] NGVIYNSMYENFSTSFWIRIPKYFRNINNEYKIISCMQNNSGWEVSLNFSNMNSKIIWTL

[1555] QDTEGIKKTVVFQYTQNINISDYINRWIFVTITNNRLSNSKIYINGRLINEESISDLGNIHAS

[1556] NNIMFKLDGCRDPHRYIWIKYFNLFDKELNKKEIKDLYDNQSNSGILKDFWGDYLQYD

[1557] KPYYMLNLYDPNKYLDVNNVGIRGYMYLKGPRGRIVTTNIYLNSTLYMGTKFIIKKYA

[1558] SGNKDNIVRNNDRVYINVVVKNKEYRLATNASQAGVEKILSAVEIPDVGNLSQVVVMK

[1559] SENDQGIRNKCKMNLQDNNGNDIGFIGFHQFNNIAKLVASNWYNRQIGKASRTFGCSW

[1560] EFIPVDDGWGESSLENLYFQGASHHHHHHHH

[1561] Nucleotide sequence (His-tag) of SEQ ID NO:31-rLC / FA

[1562] ATGCCGGTTGTGATTAACAGCTTCAATTATGATGATCCGGTGAACGATAACACCATC

[1563] ATTTATATCCGTCCGCCTTATTATGAAACCAGCAACACCTATTTCAAAGCCTTCCAG

[1564] ATTATGGATAACGTGTGGATTATTCCGGAACGTTATCGTCTGGGTATTGATCCGAGC

[1565] CTGTTTAATCCGCCTGTTAGCCTGAAAGCAGGTAGTGATGGTTATTTTGATCCGAAT

[1566] TATCTGAGCACCAACACCGAGAAAAACAAATACCTGCAGATTATGATCAAGCTGTT

[1567] CAAACGCATTAATAGCAAACCGGCAGGTCAGATTCTGCTGGAAGAAATCAAAAATG

[1568] CAATTCCGTATCTGGGCAACAGCTATACCCAAGAAGAACAGTTTACCACCAATAAT

[1569] CGTACCGTGAGCTTTAATGTTAAACTGGCCAATGGTAATATCGTTCAGCAGATGGCA

[1570] AATCTGATTATTTGGGGTCCGGGTCCTGATCTGACCACAAATAAAACCGGTGGTATC

[1571] ATCTATAGCCCGTATCAGAGCATGGAAGCAACCCCGTATAAAGATGGTTTTGGTAG

[1572] CATTATGACCGTGGAATTTAGTCCGGAATATGCAACCGCCTTTAACGATATTTCAAT

[1573] TGCAAGCCATAGTCCGTCGCTGTTTATCAAAGATCCGGCACTGATTCTGATGCATGA

[1574] ACTGATTCATGTTCTGCATGGTCTGTATGGCACCTATATTACCGAATACAAAATTAC

[1575] CCCGAATGTGGTGCAGAGCTATATGAAAGTTACCAAACCGATTACCAGCGCAGAAT

[1576] TTCTGACCTTTGGTGGTCGTGATCGCAATATTGTTCCGCAGAGCATTCAGAGCCAGC

[1577] TGTATAACAAAGTTCTGAGCGATTATAAACGTATTGCCAGCCGTCTGAATAAAGTTA

[1578] ATACCGCAACCGCACTGATCAACATCGATGAATTCAAAAACCTGTACGAGTGGAAA

[1579] TACCAGTTTGCCAAAGATAGCAATGGTGTGTATAGCGTGGATCTGAACAAATTTGA

[1580] GCAGCTGTACAAAAAAATCTATAGCTTCACCGAATTCAACCTGGCCTATGAGTTTAA

[1581] AATCAAAACCCGTCTGGGTTATCTGGCCGAAAATTTTGGTCCGTTTTATCTGCCGAA

[1582] TCTGCTGGATGATAGCATTTATACCGAAGTGGATGGTTTTAACATTGGTGCACTGAG

[1583] CATTAACTATCAGGGTCAGAATATTGGCAGCGATATCAACAGCATCAAAAAACTGC

[1584] AAGGTCAGGGTGTTGTTAGCCGTGTTGTTCGTCTGTGTAGCAATAGCGAAAATCTGT

[1585] ATTTTCAGGGTGCCAGTCATCATCACCACCATCACCATCACTGA

[1586] The polypeptide sequence (His-tag) of SEQ ID NO:32-rLC / FA

[1587] MPVVINSFNYDDPVNDNTIIYIRPPYYETSNTYFKAFQIMDNVWIIPERYRLGIDPSLFNPP

[1588] VSLKAGSDGYFDPNYLSTNTEKNKYLQIMIKLFKRINSKPAGQILLEEIKNAIPYLGNSYT

[1589] QEEQFTTNNRTVSFNVKLANGNIVQQMANLIIWGPGPDLTTNKTGGIIYSPYQSMEATP

[1590] YKDGFGSIMTVEFSPEYATAFNDISIASHSPSLFIKDPALILMHELIHVLHGLYGTYITEYK

[1591] ITPNVVQSYMKVTKPITSAEFLTFGGRDRNIVPQSIQSQLYNKVLSDYKRIASRLNKVNT

[1592] ATALINIDEFKNLYEWKYQFAKDSNGVYSVDLNKFEQLYKKIYSFTEFNLAYEFKIKTR

[1593] LGYLAENFGPFYLPNLLDDSIYTEVDGFNIGALSINYQGQNIGSDINSIKKLQGQGVVSR

[1594] VVRLCSNSENLYFQGASHHHHHHHH

[1595] Nucleotide sequence (His-tag) of SEQ ID NO:33-rBoNT / F(0)

[1596] ATGCCGGTTGTGATTAACAGCTTCAATTATAACGATCCGGTGAACGATGATACCATC

[1597] CTGTATATGCAGATTCCGTATGAAGAGAAAAGCAAAAAGTACTACAAAGCCTTTGA

[1598] GATCATGCGCAACGTTTGGATTATTCCGGAACGTAATACCATTGGCACCGATCCGA

[1599] GCGATTTTGATCCGCCTGCAAGCCTGGAAAATGGTAGCAGCGCATATTATGATCCG

[1600] AATTATCTGACCACCGATGCCGAAAAAGATCGTTATCTGAAAACCACCATCAAACT

[1601] GTTCAAACGCATTAATAGCAATCCGGCAGGCGAAGTTCTGCTGCAAGAAATTAGCT

[1602] ATGCAAAACCGTATCTGGGCAATGAACATACCCCGATTAATGAATTTCATCCGGTTA

[1603] CACGTACCACGAGCGTTAACATTAAAAGCAGCACCAATGTGAAGTCCAGCATTATT

[1604] CTGAATCTGCTGGTTTTAGGTGCAGGTCCGGATATTTTTGAAAATTCAAGCTATCCG

[1605] GTGCGCAAACTGATGGATAGCGGTGGTGTGTATGATCCGTCAAATGATGGTTTTGG

[1606] CAGCATTAACATTGTGACCTTTAGTCCGGAATATGAATACACCTTCAACGATATTAG

[1607] CGGTGGCTATAATAGCAGCACCGAAAGTTTTATTGCAGATCCGGCAATTAGCCTGG

[1608] CACACCAGCTGATTTATGCACTGCATGGTCTGTATGGTGCACGTGGTGTTACCTATA

[1609] AAGAAACCATTAAAGTTAAACAGGCACCGCTGATGATTGCGGAAAAACCGATTCGT

[1610] CTGGAAGAATTTCTGACCTTTGGTGGTCAGGATCTGAACATTATTACCAGCGCAATG

[1611] AAAGAGAAAATCTATAATAACCTGCTGGCCAACTATGAGAAAATTGCAACCCGTCT

[1612] GAGCCGTGTTAATAGCGCACCTCCTGAATATGATATCAACGAGTATAAAGACTATTT

[1613] TCAGTGGAAATACGGCCTGGATAAAAATGCAGATGGTAGCTATACCGTGAACGAGA

[1614] ACAAATTTAACGAGATCTACAAAAAACTGTATAGCTTCACCGAAATCGATCTGGCC

[1615] AACAAATTCAAAGTGAAATGCCGCAACACCTACTTCATCAAATATGGCTTTCTGAA

[1616] AGTTCCGAACCTGCTTGATGATGATATCTATACCGTTAGCGAAGGCTTTAACATTGG

[1617] TAATCTGGCCGTTAATAATCGCGGTCAGAACATTAAACTGAACCCGAAAATTATCG

[1618] ATAGCATCCCGGATAAAGGCCTGGTTGAAAAAATTGTGAAATTCTGCAAAAGCGTG

[1619] ATTCCGCGTAAAGGCACCAAAGCACCGCCTCGTCTGTGTATTCGTGTGAATAATCGT

[1620] GAACTGTTTTTTGTTGCAAGCGAGAGCAGCTATAACGAGAATGATATTAACACCCC

[1621] GAAAGAGATTGACGATACCACCAATCTGAATAACAACTATCGCAACAATCTGGATG

[1622] AAGTGATCCTGGATTATAACAGCGAAACCATTCCGCAGATTAGCAATCAGACCCTG

[1623] AATACCCTGGTTCAGGATGATAGCTATGTTCCGCGTTATGATAGCAATGGCACCAGC

[1624] GAAATTGAAGAACATAATGTGGTTGATCTGAACGTGTTCTTTTATCTGCATGCACAG

[1625] AAAGTGCCGGAAGGTGAAACCAATATTAGCCTGACCAGCAGCATTGATACCGCACT

[1626] GAGCGAAGAAAGCCAGGTTTATACCTTTTTTAGCAGCGAATTCATCAACACCATTA

[1627] ACAAACCGGTTCATGCAGCACTGTTTATTAGCTGGATTAATCAGGTGATTCGCGATT

[1628] TTACCACCGAAGCAACCCAGAAAAGCACCTTTGATAAAATTGCCGATATTAGTCTG

[1629] GTGGTGCCGTATGTTGGTCTGGCACTGAATATTGGTAATGAAGTGCAGAAAGAGAA

[1630] CTTTAAAGAAGCCTTCGAACTGTTAGGTGCCGGTATTCTGCTGGAATTTGTGCCGGA

[1631] ACTGCTGATTCCGACCATTCTGGTTTTTACCATTAAGAGCTTTATTGGCAGCAGCGA

[1632] GAACAAGAACAAAATCATTAAAGCCATCAACAACAGCCTGATGGAACGCGAAACC

[1633] AAATGGAAAGAAATTTACAGCTGGATTGTGAGCAATTGGCTGACCCGTATCAATAC

[1634] CCAGTTTAACAAACGCAAAGAACAAATGTATCAGGCCCTGCAGAATCAGGTTGATG

[1635] CAATTAAAACCGTGATCGAATACAAATACAACAACTATACCAGCGACGAACGTAAT

[1636] CGCCTGGAAAGCGAATACAACATTAATAACATTCGCGAAGAACTGAACAAAAAAG

[1637] TGAGCCTGGCAATGGAAAACATCGAACGTTTTATTACCGAAAGCAGCATCTTCTAC

[1638] CTGATGAAACTGATTAACGAAGCCAAAGTTAGCAAACTGCGCGAATATGATGAAGG

[1639] CGTTAAAGAATATCTGCTGGACTATATTAGCGAACATCGTAGCATTCTGGGTAATAG

[1640] CGTTCAAGAGCTGAATGATCTGGTTACCAGCACACTGAATAATAGCATTCCGTTTGA

[1641] ACTGAGCAGCTACACCAACGATAAAATCCTGATCCTGTACTTCAACAAACTGTACA

[1642] AGAAGATCAAGGACAACAGCATACTGGATATGCGCTATGAAAACAACAAGTTCATT

[1643] GATATCAGCGGCTATGGTAGCAACATTAGCATTAATGGTGATGTGTATATCTACAGC

[1644] ACCAACCGCAATCAGTTTGGTATTTATAGCAGCAAACCGAGCGAAGTTAATATTGC

[1645] GCAGAATAACGATATCATCTACAACGGTCGCTATCAGAACTTTAGCATTAGCTTTTG

[1646] GGTTCGCATTCCGAAATACTTTAACAAGGTGAACCTGAACAACGAGTACACCATTA

[1647] TTGATTGCATTCGCAATAATAACAGCGGCTGGAAAATCAGCCTGAACTATAACAAA

[1648] ATTATCTGGACCCTGCAGGATACCGCAGGTAATAATCAGAAACTGGTGTTTAACTA

[1649] CACCCAGATGATTAGCATCAGCGACTATATCAACAAATGGATCTTTGTGACCATTAC

[1650] CAACAATCGTCTGGGTAACAGCCGCATTTATATCAATGGCAATCTGATCGACGAAA

[1651] AAAGCATTTCAAATCTGGGCGATATTCACGTGAGCGATAACATTCTGTTCAAAATTG

[1652] TTGGCTGCAACGATACCCGTTATGTTGGTATTCGTTACTTCAAAGTGTTTGATACGG

[1653] AACTGGGCAAAACGGAAATTGAAACCCTGTATAGTGATGAACCGGATCCGAGCATT

[1654] CTGAAAGATTTTTGGGGTAATTATCTGCTGTACAACAAACGCTACTATCTGCTGAAC

[1655] CTGCTGCGTACCGATAAAAGCATTACACAGAATAGCAACTTTCTGAACATCAATCA

[1656] GCAGCGTGGTGTTTATCAGAAACCGAACATTTTTAGCAACACCCGTCTGTATACCGG

[1657] TGTGGAAGTTATTATTCGTAAAAACGGTAGCACCGATATCAGCAACACCGATAACT

[1658] TTGTGCGTAAAAATGACCTGGCCTATATTAACGTTGTTGATCGTGATGTTGAGTATC

[1659] GTCTGTATGCGGATATTAGCATTGCCAAACCGGAAAAGATTATCAAACTGATCCGT

[1660] ACCAGCAACAGCAATAATTCACTGGGTCAGATTATCGTGATGGACAGCATTGGTAA

[1661] CAATTGCACCATGAATTTCCAGAACAATAACGGTGGTAATATTGGCCTGCTGGGCTT

[1662] TCATAGCAATAATCTGGTTGCAAGCAGCTGGTATTACAACAACATCCGTAAAAATA

[1663] CCAGCAGTAATGGTTGCTTTTGGAGCTTTATCAGTAAAGAACATGGCTGGCAAGAA

[1664] AACGAGAACCTGTATTTTCAGGGTGCAAGTCATCATCACCATCACCACCATCATTAA

[1665] The polypeptide sequence (His-tag) of SEQ ID NO:34-rBoNT / F(0)

[1666] MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPERNTIGTDPSDFD

[1667] PPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRINSNPAGEVLLQEISYAKPYLGN

[1668] EHTPINEFHPPVTRTTSVNIKSSTVNKSSIILNLLVLGAGPDIHENSSYPVRKLMDSGGVYD

[1669] PSNDGFGSINIVTFSPEYETFNDISGGYNSSTESFIADPAISLAHQLIYALHGLYGARGVT

[1670] YKETIKVKQAPLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV

[1671] NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFTEIDLANKFKV

[1672] KCRNTYFIKYGFLKVPNLLDDDIYTVSEGFNIGNLAVNNRGQNIKLNPKIIDSIPDKGLVE

[1673] KIVKFCKSVIPRKGTKAPPRLCIRVNNRELFFVASESSYNDINTKEIDDTTTNLNNYR

[1674] NNLDEVILDYNSETIPQISNQTLNTLVQDDSYVPRYDSNGTSEIEEHNVVDLNVFFYLHA

[1675] QKVPEGETNISLTSSIDTALSEESQVYTFFSSEFINTINKPVHAALFISWINQVIRDFTTEAT

[1676] QKSTFDKIADISLVVPYVGLALNIGNEVQKENFKEAFELLGAGILLEFVPELLIPTILVFTI

[1677] KSFIGSSENKNKIIKAINNSLMERETKWKEIYSWIVSNWLTRINTQFNRKKEQMYQALQ

[1678] NQVDAIKTVIEYKYNNYTSDERNRLESEYNINNIREELNKKVSLAMENIERFITESSIFYL

[1679] MKLINEAKVSKLREYDEGVKEYLLDYISEHRSILGNSVQELNDLVTSTLNNSIPFELSSYT

[1680] NDKILILYFNKLYKKIKDNSILDMRYENNKFIDISGYGSNISINGDVYIYSTNRNQFGIYSS

[1681] KPSEVNIAQNNDIIYNGRYQNFSISFWVRIPKYFNKVNLNNEYTIIDCIRNNNSGWKISLN

[1682] YNKIIWTLQDTAGNNQKLVFNYTQMISISDYINKWIFVTITNNRLGNSRIYINGNLIDEKS

[1683] ISNLGDIHVSDNILFKIVGCNDTRYVGIRYFKVFDTELGKTEIETLYSDEPDPSILKDFWG

[1684] NYLLYNKRYYLLNLLRTDKSITQNSNFLNINQQRGVYQKPNIFSNTRLYTGVEVIIRKNG

[1685] STDISNTDNFVRKNDLAYINVVDRDVEYRLYADISIAKPEKIIKLIRTSNSNNSLGQIIVM

[1686] DSIGNNCTMNFQNNNGGNIGLLGFHSNNLVASSWYYNNIRKNTSSNGCFWSFISKEHG

[1687] WQENENLYFQGASHHHHHHHH

[1688] SEQ ID NO:35-rL H N / F nucleotide sequence (His-tag)

[1689] ATGCCGGTTGTGATTAACAGCTTCAATTATAACGATCCGGTGAACGATGATACCATC

[1690] CTGTATATGCAGATTCCGTATGAAGAGAAAAGCAAAAAGTACTACAAAGCCTTTGA

[1691] GATCATGCGCAACGTTTGGATTATTCCGGAACGTAATACCATTGGCACCGATCCGA

[1692] GCGATTTTGATCCGCCTGCAAGCCTGGAAAATGGTAGCAGCGCATATTATGATCCG

[1693] AATTATCTGACCACCGATGCCGAAAAAGATCGTTATCTGAAAACCACCATCAAACT

[1694] GTTCAAACGCATTAATAGCAATCCGGCAGGCGAAGTTCTGCTGCAAGAAATTAGCT

[1695] ATGCAAAACCGTATCTGGGCAATGAACATACCCCGATTAATGAATTTCATCCGGTTA

[1696] CACGTACCACGAGCGTTAACATTAAAAGCAGCACCAATGTGAAGTCCAGCATTATT

[1697] CTGAATCTGCTGGTTTTAGGTGCAGGTCCGGATATTTTTGAAAATTCAAGCTATCCG

[1698] GTGCGCAAACTGATGGATAGCGGTGGTGTGTATGATCCGTCAAATGATGGTTTTGG

[1699] CAGCATTAACATTGTGACCTTTAGTCCGGAATATGAATACACCTTCAACGATATTAG

[1700] CGGTGGCTATAATAGCAGCACCGAAAGTTTTATTGCAGATCCGGCAATTAGCCTGG

[1701] CACATGAACTGATTCATGCACTGCATGGTCTGTATGGTGCACGTGGTGTTACCTATA

[1702] AAGAAACCATTAAAGTTAAACAGGCACCGCTGATGATTGCGGAAAAACCGATTCGT

[1703] CTGGAAGAATTTCTGACCTTTGGTGGTCAGGATCTGAACATTATTACCAGCGCAATG

[1704] AAAGAGAAAATCTATAATAACCTGCTGGCCAACTATGAGAAAATTGCAACCCGTCT

[1705] GAGCCGTGTTAATAGCGCACCTCCTGAATATGATATCAACGAGTATAAAGACTATTT

[1706] TCAGTGGAAATACGGCCTGGATAAAAATGCAGATGGTAGCTATACCGTGAACGAGA

[1707] ACAAATTTAACGAGATCTACAAAAAACTGTATAGCTTCACCGAAATCGATCTGGCC

[1708] AACAAATTCAAAGTGAAATGCCGCAACACCTACTTCATCAAATATGGCTTTCTGAA

[1709] AGTTCCGAACCTGCTTGATGATGATATCTATACCGTTAGCGAAGGCTTTAACATTGG

[1710] TAATCTGGCCGTTAATAATCGCGGTCAGAACATTAAACTGAACCCGAAAATTATCG

[1711] ATAGCATCCCGGATAAAGGCCTGGTTGAAAAAATTGTGAAATTCTGCAAAAGCGTG

[1712] ATTCCGCGTAAAGGCACCAAAGCACCGCCTCGTCTGTGTATTCGTGTGAATAATCGT

[1713] GAACTGTTTTTTGTTGCAAGCGAGAGCAGCTATAACGAGAATGATATTAACACCCC

[1714] GAAAGAGATTGACGATACCACCAATCTGAATAACAACTATCGCAACAATCTGGATG

[1715] AAGTGATCCTGGATTATAACAGCGAAACCATTCCGCAGATTAGCAATCAGACCCTG

[1716] AATACCCTGGTTCAGGATGATAGCTATGTTCCGCGTTATGATAGCAATGGCACCAGC

[1717] GAAATTGAAGAACATAATGTGGTTGATCTGAACGTGTTCTTTTATCTGCATGCACAG

[1718] AAAGTGCCGGAAGGTGAAACCAATATTAGCCTGACCAGCAGCATTGATACCGCACT

[1719] GAGCGAAGAAAGCCAGGTTTATACCTTTTTTAGCAGCGAATTCATCAACACCATTA

[1720] ACAAACCGGTTCATGCAGCACTGTTTATTAGCTGGATTAATCAGGTGATTCGCGATT

[1721] TTACCACCGAAGCAACCCAGAAAAGCACCTTTGATAAAATTGCCGATATTAGTCTG

[1722] GTGGTGCCGTATGTTGGTCTGGCACTGAATATTGGTAATGAAGTGCAGAAAGAGAA

[1723] CTTTAAAGAAGCCTTCGAACTGTTAGGTGCCGGTATTCTGCTGGAATTTGTGCCGGA

[1724] ACTGCTGATTCCGACCATTCTGGTTTTTACCATTAAGAGCTTTATTGGCAGCAGCGA

[1725] GAACAAGAACAAAATCATTAAAGCCATCAACAACAGCCTGATGGAACGCGAAACC

[1726] AAATGGAAAGAAATTTACAGCTGGATTGTGAGCAATTGGCTGACCCGTATCAATAC

[1727] CCAGTTTAACAAACGCAAAGAACAAATGTATCAGGCCCTGCAGAATCAGGTTGATG

[1728] CAATTAAAACCGTGATCGAATACAAATACAACAACTATACCAGCGACGAACGTAAT

[1729] CGCCTGGAAAGCGAATACAACATTAATAACATTCGCGAAGAACTGAACAAAAAAG

[1730] TGAGCCTGGCAATGGAAAACATCGAACGTTTTATTACCGAAAGCAGCATCTTCTAC

[1731] CTGATGAAACTGATTAACGAAGCCAAAGTTAGCAAACTGCGCGAATATGATGAAGG

[1732] CGTTAAAGAATATCTGCTGGACTATATTAGCGAACATCGTAGCATTCTGGGTAATAG

[1733] CGTTCAAGAGCTGAATGATCTGGTTACCAGCACACTGAATAATAGCATTCCGTTTGA

[1734] ACTGAGCAGCTACACCAACGATAAAATCCTGATCCTGTACTTCAACAAACTGTACA

[1735] AGAAAGAAAACCTGTATTTTCAGGGTGCAAGCCATCATCACCACCATCACCATCAT

[1736] TAA

[1737] SEQ ID NO:36-rL H N / F polypeptide sequence (His-tag)

[1738] MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPERNTIGTDPSDFD

[1739] PPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRINSNPAGEVLLQEISYAKPYLGN

[1740] EHTPINEFHPPVTRTTSVNIKSSTVNKSSIILNLLVLGAGPDIHENSSYPVRKLMDSGGVYD

[1741] PSNDGFGSINIVTFSPEYETFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGARGVT

[1742] YKETIKVKQAPLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV

[1743] NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFTEIDLANKFKV

[1744] KCRNTYFIKYGFLKVPNLLDDDIYTVSEGFNIGNLAVNNRGQNIKLNPKIIDSIPDKGLVE

[1745] KIVKFCKSVIPRKGTKAPPRLCIRVNNRELFFVASESSYNDINTKEIDDTTTNLNNYR

[1746] NNLDEVILDYNSETIPQISNQTLNTLVQDDSYVPRYDSNGTSEIEEHNVVDLNVFFYLHA

[1747] QKVPEGETNISLTSSIDTALSEESQVYTFFSSEFINTINKPVHAALFISWINQVIRDFTTEAT

[1748] QKSTFDKIADISLVVPYVGLALNIGNEVQKENFKEAFELLGAGILLEFVPELLIPTILVFTI

[1749] KSFIGSSENKNKIIKAINNSLMERETKWKEIYSWIVSNWLTRINTQFNRKKEQMYQALQ

[1750] NQVDAIKTVIEYKYNNYTSDERNRLESEYNINNIREELNKKVSLAMENIERFITESSIFYL

[1751] MKLINEAKVSKLREYDEGVKEYLLDYISEHRSILGNSVQELNDLVTSTLNNSIPFELSSYT

[1752] NDKILILYFNKLYKKENLYFQGASHHHHHHH

[1753] SEQ ID NO:37-rH C / F nucleotide sequence (His-tag)

[1754] ATGATCAAGGATAACAGCATTCTGGATATGCGCTATGAGAACAACAAATTCATTGA

[1755] TATTAGCGGCTATGGCAGCAACATTAGCATTAATGGTGATGTGTATATCTACAGCAC

[1756] CAACCGTAATCAGTTTGGCATTTATAGCAGCAAACCGAGCGAAGTTAATATTGCCC

[1757] AGAACAACGATATCATCTATAACGGTCGCTATCAGAACTTCAGCATTAGCTTTTGGG

[1758] TTCGCATTCCGAAATACTTCAATAAGGTGAACCTGAACAACGAGTATACCATCATTG

[1759] ATTGCATTCGCAATAATAACAGCGGCTGGAAAATTAGCCTGACTACAACAAAATT

[1760] ATCTGGACCCTGCAGGATACCGCAGGTAAATATCAGAAACTGGTGTTTAACTACAC

[1761] CCAGATGATTAGCATCAGCGACTATATCAACAAATGGATCTTTGTGACCATTACCAA

[1762] TAATCGCCTGGGTAATAGCCGCATTTATATCAATGGTAACCTGATCGATGAGAAAA

[1763] GCATTAGCAATCTGGGTGATATTCATGTGAGCGATAACATCCTGTTTAAAATCGTGGG

[1764] GTTGTAACGATACCCGTTATGTTGGTATTCGCTACTTCAAAGTGTTTGATACCGAAC

[1765] TGGGTAAAACCGAAATTGAACCCTGTATAGTGATGAACCGGATCCGAGCATTCTG

[1766] AAAGATTTTTGGGGTAATTATCTGCTGTACAACAAACGCTACTATCTGCTGAATCTG

[1767] CTGCGTACCGATAAATCAATTACCCAGAATAGCAACTTCCTGAACATTAATCAGCA

[1768] GCGTGGTGTTTATCAGAAACCGAACATTTTTAGCAACACCCGTCTGTATACCGGTGT

[1769] GGAAGTTATTATTCGTAAAAAATGGCAGCACCGATATCAGCAAACCGATAACTTTG

[1770] TTCGCAAAAATGATCTGGCGTATATCAACGTTGTTGATCGTGATGTTGAATATCGTC

[1771] TGTATGCCGATATTAGCATTGCCAAACCGGAAAAAATCATCAAACTGATCCGTACC

[1772] AGCAACAGCAATAATTCACTGGGTCAGATTATTGTGATGGATAGCATTGGTAATAA

[1773] CTGCACCATGAACTTTCAGAAACATAACGGTGGTAATATTGGTCTGCTGGCTTTCA

[1774] TAGTAATAATCTGGTTGCAAGCAGCTGGTATTATAACAACATCCGTAAAAATACCA

[1775] GCAGCAATGGTTGCTTTTGGAGCTTTATTAGCAAAGAACATGGCTGGCAAGAAAAC

[1776] GAGAATCTGTATTTTCAGGGTGCAAGTCATCATCACCACCATCACCATCATTAA

[1777] SEQ ID NO:38-rH C / F polypeptide sequence (His-tag)

[1778] MIKDNSILDMRYENNKFIDISGYGSNISINGDVYIYSTNRNQFGIYSSKPSEVNIAQNNDII

[1779] YNGRYQNFSISFWVRIPKYFNKVNLNNEYTIIDCIRNNNSGWKISLNYNKIIWTLQDTAG

[1780] NNQKLVFNYTQMISISDYINKWIFVTITNNRLGNSRIYINGNLIDEKSISNLGDIHVSDNIL

[1781] FKIVGCNDTRYVGIRYFKVFDTELGKTEIETLYSDEPDPSILKDFWGNYLLYNKRYYLLN

[1782] LLRTDKSITQNSNFLNINQQRGVYQKPNIFSNTRLYTGVEVIIRKNGSTDISNTDNFVRKN

[1783] DLAYINVVDRDVEYRLYADISIAKPEKIIKLIRTSNSNNSLGQIIVMDSIGNNCTMNFQNN

[1784] NGGNIGLLGFHSNNLVASSWYYNNIRKNTSSNGCFWSFISKEHGWQENENLYFQGASH

[1785] HHHHHHH

[1786] Nucleotide sequence (His-tag) of SEQ ID NO:39-rLC / F

[1787] ATGCCGGTTGTGATTAACAGCTTCAATTATAACGATCCGGTGAACGATGATACCATC

[1788] CTGTATATGCAGATTCCGTATGAAGAGAAAAGCAAAAAGTACTACAAAGCCTTTGA

[1789] GATCATGCGCAACGTTTGGATTATTCCGGAACGTAATACCATTGGCACCGATCCGA

[1790] GCGATTTTGATCCGCCTGCAAGCCTGGAAAATGGTAGCAGCGCATATTATGATCCG

[1791] AATTATCTGACCACCGATGCCGAAAAAGATCGTTATCTGAAAACCACCATCAAACT

[1792] GTTCAAACGCATTAATAGCAATCCGGCAGGCGAAGTTCTGCTGCAAGAAATTAGCT

[1793] ATGCAAAACCGTATCTGGGCAATGAACATACCCCGATTAATGAATTTCATCCGGTTA

[1794] CACGTACCACGAGCGTTAACATTAAAAGCAGCACCAATGTGAAGTCCAGCATTATT

[1795] CTGAATCTGCTGGTTTTAGGTGCAGGTCCGGATATTTTTGAAAATTCAAGCTATCCG

[1796] GTGCGCAAACTGATGGATAGCGGTGGTGTGTATGATCCGTCAAATGATGGTTTTGG

[1797] CAGCATTAACATTGTGACCTTTAGTCCGGAATATGAATACACCTTCAACGATATTAG

[1798] CGGTGGCTATAATAGCAGCACCGAAAGTTTTATTGCAGATCCGGCAATTAGCCTGG

[1799] CACATGAACTGATTCATGCACTGCATGGTCTGTATGGTGCACGTGGTGTTACCTATA

[1800] AAGAAACCATTAAAGTTAAACAGGCACCGCTGATGATTGCGGAAAAACCGATTCGT

[1801] CTGGAAGAATTTCTGACCTTTGGTGGTCAGGATCTGAACATTATTACCAGCGCAATG

[1802] AAAGAGAAAATCTATAATAACCTGCTGGCCAACTATGAGAAAATTGCAACCCGTCT

[1803] GAGCCGTGTTAATAGCGCACCTCCTGAATATGATATCAACGAGTATAAAGACTATTT

[1804] TCAGTGGAAATACGGCCTGGATAAAAATGCAGATGGTAGCTATACCGTGAACGAGA

[1805] ACAAATTTAACGAGATCTACAAAAAACTGTATAGCTTCACCGAAATCGATCTGGCC

[1806] AACAAATTCAAAGTGAAATGCCGCAACACCTACTTCATCAAATATGGCTTTCTGAA

[1807] AGTTCCGAACCTGCTTGATGATGATATCTATACCGTTAGCGAAGGCTTTAACATTGG

[1808] TAATCTGGCCGTTAATAATCGCGGTCAGAACATTAAACTGAACCCGAAAATTATCG

[1809] ATAGCATCCCGGATAAAGGCCTGGTTGAAAAAATTGTGAAATTCTGCAAAAGCGAG

[1810] AACCTGTATTTTCAGGGTGCAAGTCATCATCACCATCACCACCATCATTAA

[1811] The polypeptide sequence (His-tag) of SEQ ID NO:40-rLC / F

[1812] MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPERNTIGTDPSDFD

[1813] PPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRINSNPAGEVLLQEISYAKPYLGN

[1814] EHTPINEFHPVTRTTSVNIKSSTNVKSSIILNLLVLGAGPDIFENSSYPVRKLMDSGGVYD

[1815] PSNDGFGSINIVTFSPEYEYTFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGARGVT

[1816] YKETIKVKQAPLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV

[1817] NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFTEIDLANKFKV

[1818] KCRNTYFIKYGFLKVPNLLDDDIYTVSEGFNIGNLAVNNRGQNIKLNPKIIDSIPDKGLVE

[1819] KIVKFCKSENLYFQGASHHHHHHHH

[1820] SEQ ID NO:41 - rH of cations C The nucleotide sequence of / A (His-tag)

[1821] ATGATCATCAACACCAGCATTCTGAACCTGCGTTATGAAAGCAAACATCTGATTGAT

[1822] CTGAGCCGTTATGCCAGCAAAATCAATATAGGCAGCAAGGTTAACTTCGACCCGAT

[1823] TGACAAAAATCAGATACAGCTGTTTAATCTGGAAAGCAGCAAAATTGAGGTGATCC

[1824] TGAAAAAAGCGATCGTGTATAATAGCATGTACGAGAATTTTTCGACCAGCTTTTGG

[1825] ATTCGCATCCCGAAATACTTTAACAAGATTAGCCTGAACAACGAGTATACCATCATT

[1826] AACTGCATGGAAAACAATAGCGGTTGGAAAGTCAGCCTGAATTATGGCGAAATTAT

[1827] CTGGACCCTGCAGGATACCAAAGAAATCAAACAGCGTGTGGTGTTCAAATACAGCC

[1828] AGATGATTAATATCAGCGACTATATCAACCGCTGGATTTTTGTGACCATTACCAATA

[1829] ATCGGCTGAACAAGAGCAAGATCTATATTAACGGTCGTCTGATTGACCAGAAACCG

[1830] ATTAGTAATCTGGGTAATATTCATGCGAGCAACAAAATCATGTTTAAACTGGATGGT

[1831] TGCCGTGATACCCATCGTTATATTTGGATCAAATACTTCAACCTGTTCGATAAAGAG

[1832] TTGAACGAAAAAGAAATTAAAGACCTGTACGATAACCAGAGCAATAGCGGCATACT

[1833] GAAAGATTTTTGGGGAGATTATCTGCAGTATGACAAACCGTATTATATGCTGAATCT

[1834] GTACGACCCGAATAAATACGTGGATGTTAATAATGTGGGCATCCGTGGTTATATGT

[1835] ACCTGAAAGGTCCGCGTGGTAGCGTTATGACCACAAACATTTATCTGAATAGCAGC

[1836] CTGTATCGCGGAACCAAATTCATCATTAAAAAGTATGCCAGCGGCAACAAGGATAA

[1837] TATTGTGCGTAATAATGATCGCGTGTACATTAACGTTGTGGTGAAGAATAAAGAAT

[1838] ATCGCCTGGCAACCAATGCAAGCCAGGCAGGCGTTGAAAAAATTCTGAGTGCCCTG

[1839] GAAATTCCGGATGTTGGTAATCTGAGCCAGGTTGTTGTGATGAAAAGCAAAAACGA

[1840] TAAAGGCATCACCAACAAATGCAAGATGAATCTGCAGGACAATAACGGCAATGAT

[1841] ATTGGCTTCATTGGCTTTCACCAGTTTAACAACATTGCAAAACTGGTTGCGAGCAAT

[1842] TGGTATAATCGTCAGATTGAACGTAGCAGTCGTACCCTGGGTTGTAGCTGGGAATTT

[1843] ATCCCTGTGGATGATGGTTGGGGTGAACGTCCGCTGAAGCTTGCGGCCGCACTCGA

[1844] GCACCACCACCACCACCACTGA

[1845] SEQ ID NO:42 - rH of cations C / A polypeptide sequence (His-tag)

[1846] MIINTSILNLRYESKHLIDLSRYASKINIGSKVNFDPIDKNQIQLFNLESSKIEVILKKAIVY

[1847] NSMYENFSTSFWIRIPKYFNKISLNNEYTIINCMENNSGWKVSLNYGEIIWTLQDTKEIK

[1848] QRVVFKYSQMINISDYINRWIFVTITNNRLNKSKIYINGRLIDQKPISNLGNIHASNKIMFK

[1849] LDGCRDTHRYIWIKYFNLFDKELNEKEIKDLYDNQSNSGILKDFWGDYLQYDKPYYML

[1850] NLYDPNKYVDVNNVGIRGYMYLKGPRGSVMTTNIYLNSSLYRGTKFIIKKYASGNKDN

[1851] IVRNNDRVYINVVVKNKEYRLATNASQAGVEKILSALEIPDVGNLSQVVVMKSKNDKG

[1852] ITNKCKMNLQDNNGNDIGFIGFHQFNNIAKLVASNWYNRQIERSSRTLGCSWEFIPVDD

[1853] GWGERPLKLAAALEHHHHHH

[1854] SEQ ID NO:43-rH C The nucleotide sequence of / AB (His-tag)

[1855] ATGATTCTGAACAATATTATCCTGAACCTGCGTTACAAAGACAACAATCTGATCGAT

[1856] CTGAGCGGCTATGGTGCAAAAGTTGAAGTCTACGACGGTGTCGAACTGAACGATAA

[1857] AAACCAGTTCAAACTGACCTCATCGGCTAACTCAAAAATTCGTGTGACGCAGAACC

[1858] AAAACATCATCTTCAACTCGGTCTTTCTGGACTTCAGCGTGTCTTTCTGGATTCGCAT

[1859] CCCGAAATATAAAAATGATGGCATCCAGAACTACATCCATAACGAATACACCATCA

[1860] TCAACTGTATGAAAAACAACAGTGGTTGGAAAATTTCCATCCGTGGCAACCGCATT

[1861] ATCTGGACCCTGATTGATATCAATGGTAAAACGAAAAGCGTGTTTTTCGAATACAA

[1862] CATCCGTGAAGATATCTCTGAATACATCAATCGCTGGTTTTTCGTGACCATTACGAA

[1863] CAATCTGAACAATGCGAAAATCTATATCAACGGCAAACTGGAAAGTAATACCGACA

[1864] TCAAAGATATTCGTGAAGTTATCGCCAACGGTGAAATCATCTTCAAACTGGATGGC

[1865] GACATCGATCGCACCCAGTTCATTTGGATGAAATACTTCTCCATCTTCAACACGGAA

[1866] CTGAGTCAGTCCAATATCGAAGAACGCTACAAAATCCAATCATACTCGGAATACCT

[1867] GAAAGATTTCTGGGGTAACCCGCTGATGTACAACAAAGAATACTACATGTTCAACG

[1868] CGGGCAACAAAAACTCATACATCAAACTGAAAAAAGATTCGCCGGTGGGTGAAATC

[1869] CTGACCCGTAGCAAATACAACCAGAACTCTAAATACATCAACTATCGCGATCTGTA

[1870] CATTGGCGAAAAATTTATTATCCGTCGCAAAAGCAACTCTCAGAGTATTAATGATG

[1871] ACATCGTGCGTAAAGAAGACTACATCTATCTGGATTTCTTTAATCTGAACCAAGAAT

[1872] GGCGCGTTTATACCTACAAATACTTCAAAAAAGAAGAAATGAAACTGTTCCTGGCC

[1873] CCGATTTACGACAGCGATGAATTTTACAACACCATCCAGATCAAAGAATACGATGA

[1874] ACAGCCGACGTATAGTTGCCAACTGCTGTTCAAAAAAGACGAAGAATCCACCGATG

[1875] AAATTGGCCTGATTGGTATCCACCGTTTCTATGAAAGCGGTATCGTTTTCGAAGAAT

[1876] ACAAAGATTACTTCTGTATCTCTAAATGGTATCTGAAAGAAGTCAAACGCAAACCG

[1877] TACAACCTGAAACTGGGCTGCAACTGGCAATTTATCCCGAAAGACGAAGGCTGGAC

[1878] CGAAAAGCTTGCGGCCGCACTCGAGCACCACCACCACCACCACTGA

[1879] SEQ ID NO:44-rH C / AB polypeptide sequence (His-tag)

[1880] MILNNIILNLRYKDNNLIDLSGYGAKVEVYDGVELNDKNQFKLTSSANSKIRVTQNQNII

[1881] FNSVFLDFSVSFWIRIPKYKNDGIQNYIHNEYTIINCMKNNSGWKISIRGNRIIWTLIDING

[1882] KTKSVFFEYNIREDISEYINRWFFVTITNNLNNAKIYINGKLESNTDIKDIREVIANGEIIFK

[1883] LDGDIDRTQFIWMKYFSIFNTELSQSNIEERYKIQSYSEYLKDFWGNPLMYNKEYYMFN

[1884] AGNKNSYIKLKKDSPVGEILTRSKYNQNSKYINYRDLYIGEKFIIRRKSNSQSINDDIVRK

[1885] EDYIYLDFFNLNQEWRVYTYKYFKKEEMKLFLAPIYDSDEFYNTIQIKEYDEQPTYSCQ

[1886] LLFKKDEESTDEIGLIGIHRFYESGIVFEEYKDYFCISKWYLKEVKRKPYNLKLGCNWQF

[1887] IPKDEGWTEKLAAALEHHHHHH

[1888] SEQ ID NO:45-rH C The nucleotide sequence (His-tag) of variant Y1117VH1253K.

[1889] ATGATCATCAATACTAGCATTCTGAACCTGCGTTACGAGAGCAATCATCTGATTGAT

[1890] CTGAGCCGTTATGCAAGCAAGATCAACATCGGTAGCAAGGTCAATTTTGACCCGAT

[1891] CGATAAGAACCAGATCCAGCTGTTTAATCTGGAATCGAGCAAAATTGAGGTTATCC

[1892] TGAAAAACGCCATTGTCTACAACTCCATGTACGAGAATTTCTCCACCAGCTTCTGGA

[1893] TTCGCATCCCGAAATACTTCAACAGCATTAGCCTGAACAACGAGTATACTATCATCA

[1894] ACTGTATGGAGAACAACAGCGGTTGGAAGGTGTCTCTGAACTATGGTGAGATCATT

[1895] TGGACCTTGCAGGACACCCAAGAGATCAAGCAGCGCGTCGTGTTCAAGTACTCTCA

[1896] AATGATCAACATTTCCGATTACATTAATCGTTGGATCTTCGTGACCATTACGAATAA

[1897] CCGTCTGAATAACAGCAAGATTTACATCAATGGTCGCTTGATCGATCAGAAACCGA

[1898] TTAGCAACCTGGGTAATATCCACGCAAGCAACAACATTATGTTCAAATTGGACGGT

[1899] TGCCGCGATACCCATCGTTATATCTGGATCAAGTATTTCAACCTGTTTGATAAAGAA

[1900] CTGAATGAGAAGGAGATCAAAGATTTGTATGACAACCAATCTAACAGCGGCATTTT

[1901] GAAGGACTTCTGGGGCGATTATCTGCAATACGATAAGCCGTACTATATGCTGAACC

[1902] TGgtTGATCCGAACAAATATGTGGATGTCAATAATGTGGGTATTCGTGGTTACATGTA

[1903] TTTGAAGGGTCCGCGTGGCAGCGTTATGACGACCAACATTTACCTGAACTCTAGCCT

[1904] GTACCGTGGTACGAAATTCATCATTAAGAAATATGCCAGCGGCAACAAAGATAACA

[1905] TTGTGCGTAATAACGATCGTGTCTACATCAACGTGGTCGTGAAGAATAAAGAGTAC

[1906] CGTCTGGCGACCAACGCTTCGCAGGCGGGTGTTGAGAAAATTCTGAGCGCGTTGGA

[1907] GATCCCTGATGTCGGTAATCTGAGCCAAGTCGTGGTTATGAAGAGCAAGAACGACC

[1908] AGGGTATCACTAACAAGTGCAAGATGAACCTGCAAGACAACAATGGTAACGACATC

[1909] GGCTTTATTGGTTTCaAaCAGTTCAACAATATTGCTAAACTGGTAGCGAGCAATTGGT

[1910] ACAATCGTCAGATTGAGCGCAGCAGCCGTACTTTGGGCTGTAGCTGGGAGTTTATCC

[1911] CGGTCGATGATGGTTGGGGCGAACGTCCGCTGCACCATCACCATCACCATCACCAT

[1912] CACCATT

[1913] SEQ ID NO:46-rH C / A variant Y1117VH1253K polypeptide sequence (His-tag)

[1914] MIINTSILNLRYESNHLIDLSRYASKINIGSKVNFDPIDKNQIQLFNLESSKIEVILKNAIVY

[1915] NSMYENFSTSFWIRIPKYFNSISLNNEYTIINCMENNSGWKVSLNYGEIIWTLQDTQEIKQ

[1916] RVVFKYSQMINISDYINRWIFVTITNNRLNNSKIYINGRLIDQKPISNLGNIHASNNIMFKL

[1917] DGCRDTHRYIWIKYFNLFDKELNEKEIKDLYDNQSNSGILKDFWGDYLQYDKPYYMLN

[1918] LVDPNKYVDVNNVGIRGYMYLKGPRGSVMTTNIYLNSSLYRGTKFIIKKYASGNKDNI

[1919] VRNNDRVYINVVVKNKEYRLATNASQAGVEKILSALEIPDVGNLSQVVVMKSKNDQGI

[1920] TNKCKMNLQDNNGNDIGFIGFKQFNNIAKLVASNWYNRQIERSSRTLGCSWEFIPVDDG

[1921] WGERPLHHHHHHHHHH

[1922] SEQ ID NO:47-rH C The nucleotide sequence of variant Y1117V F1252Y H1253KL1278F (His-marked) sign)

[1923] ATGATCATCAATACTAGCATTCTGAACCTGCGTTACGAGAGCAATCATCTGATTGAT

[1924] CTGAGCCGTTATGCAAGCAAGATCAACATCGGTAGCAAGGTCAATTTTGACCCGAT

[1925] CGATAAGAACCAGATCCAGCTGTTTAATCTGGAATCGAGCAAAATTGAGGTTATCC

[1926] TGAAAAACGCCATTGTCTACAACTCCATGTACGAGAATTTCTCCACCAGCTTCTGGA

[1927] TTCGCATCCCGAAATACTTCAACAGCATTAGCCTGAACAACGAGTATACTATCATCA

[1928] ACTGTATGGAGAACAACAGCGGTTGGAAGGTGTCTCTGAACTATGGTGAGATCATT

[1929] TGGACCTTGCAGGACACCCAAGAGATCAAGCAGCGCGTCGTGTTCAAGTACTCTCA

[1930] AATGATCAACATTTCCGATTACATTAATCGTTGGATCTTCGTGACCATTACGAATAA

[1931] CCGTCTGAATAACAGCAAGATTTACATCAATGGTCGCTTGATCGATCAGAAACCGA

[1932] TTAGCAACCTGGGTAATATCCACGCAAGCAACAACATTATGTTCAAATTGGACGGT

[1933] TGCCGCGATACCCATCGTTATATCTGGATCAAGTATTTCAACCTGTTTGATAAAGAA

[1934] CTGAATGAGAAGGAGATCAAAGATTTGTATGACAACCAATCTAACAGCGGCATTTT

[1935] GAAGGACTTCTGGGGCGATTATCTGCAATACGATAAGCCGTACTATATGCTGAACC

[1936] TGgtTGATCCGAACAAATATGTGGATGTCAATAATGTGGGTATTCGTGGTTACATGTA

[1937] TTTGAAGGGTCCGCGTGGCAGCGTTATGACGACCAACATTTACCTGAACTCTAGCCT

[1938] GTACCGTGGTACGAAATTCATCATTAAGAAATATGCCAGCGGCAACAAAGATAACA

[1939] TTGTGCGTAATAACGATCGTGTCTACATCAACGTGGTCGTGAAGAATAAAGAGTAC

[1940] CGTCTGGCGACCAACGCTTCGCAGGCGGGTGTTGAGAAAATTCTGAGCGCGTTGGA

[1941] GATCCCTGATGTCGGTAATCTGAGCCAAGTCGTGGTTATGAAGAGCAAGAACGACC

[1942] AGGGTATCACTAACAAGTGCAAGATGAACCTGCAAGACAACAATGGTAACGACATC

[1943] GGCTTTATTGGTTaCaAaCAGTTCAACAATATTGCTAAACTGGTAGCGAGCAATTGGT

[1944] ACAATCGTCAGATTGAGCGCAGCAGCCGTACTTTtGGCTGTAGCTGGGAGTTTATCC

[1945] CGGTCGATGATGGTTGGGGCGAACGTCCGCTGCACCATCACCATCACCATCACCAT

[1946] CACCATTAA

[1947] SEQ ID NO:48-rH C / A Peptide sequence (His-tag) of variant Y1117V F1252YH1253KL1278F

[1948] MIINTSILNLRYESNHLIDLSRYASKINIGSKVNFDPIDKNQIQLFNLESSKIEVILKNAIVY

[1949] NSMYENFSTSFWIRIPKYFNSISLNNEYTIINCMENNSGWKVSLNYGEIIWTLQDTQEIKQ

[1950] RVVFKYSQMINISDYINRWIFVTITNNRLNNSKIYINGRLIDQKPISNLGNIHASNNIMFKL

[1951] DGCRDTHRYIWIKYFNLFDKELNEKEIKDLYDNQSNSGILKDFWGDYLQYDKPYYMLN

[1952] LVDPNKYVDVNNVGIRGYMYLKGPRGSVMTTNIYLNSSLYRGTKFIIKKYASGNKDNI

[1953] VRNNDRVYINVVVKNKEYRLATNASQAGVEKILSALEIPDVGNLSQVVVMKSKNDQGI

[1954] TNKCKMNLQDNNGNDIGFIGYKQFNNIAKLVASNWYNRQIERSSRTFGCSWEFIPVDD

[1955] GWGERPLHHHHHHHHHH

[1956] SEQ ID NO:49-rH C The nucleotide sequence (His-tag) of variant A Y1117V F1252YH1253KL1278H.

[1957] ATGATCATCAATACTAGCATTCTGAACCTGCGTTACGAGAGCAATCATCTGATTGAT

[1958] CTGAGCCGTTATGCAAGCAAGATCAACATCGGTAGCAAGGTCAATTTTGACCCGAT

[1959] CGATAAGAACCAGATCCAGCTGTTTAATCTGGAATCGAGCAAAATTGAGGTTATCC

[1960] TGAAAAACGCCATTGTCTACAACTCCATGTACGAGAATTTCTCCACCAGCTTCTGGA

[1961] TTCGCATCCCGAAATACTTCAACAGCATTAGCCTGAACAACGAGTATACTATCATCA

[1962] ACTGTATGGAGAACAACAGCGGTTGGAAGGTGTCTCTGAACTATGGTGAGATCATT

[1963] TGGACCTTGCAGGACACCCAAGAGATCAAGCAGCGCGTCGTGTTCAAGTACTCTCA

[1964] AATGATCAACATTTCCGATTACATTAATCGTTGGATCTTCGTGACCATTACGAATAA

[1965] CCGTCTGAATAACAGCAAGATTTACATCAATGGTCGCTTGATCGATCAGAAACCGA

[1966] TTAGCAACCTGGGTAATATCCACGCAAGCAACAACATTATGTTCAAATTGGACGGT

[1967] TGCCGCGATACCCATCGTTATATCTGGATCAAGTATTTCAACCTGTTTGATAAAGAA

[1968] CTGAATGAGAAGGAGATCAAAGATTTGTATGACAACCAATCTAACAGCGGCATTTT

[1969] GAAGGACTTCTGGGGCGATTATCTGCAATACGATAAGCCGTACTATATGCTGAACC

[1970] TGgtTGATCCGAACAAATATGTGGATGTCAATAATGTGGGTATTCGTGGTTACATGTA

[1971] TTTGAAGGGTCCGCGTGGCAGCGTTATGACGACCAACATTTACCTGAACTCTAGCCT

[1972] GTACCGTGGTACGAAATTCATCATTAAGAAATATGCCAGCGGCAACAAAGATAACA

[1973] TTGTGCGTAATAACGATCGTGTCTACATCAACGTGGTCGTGAAGAATAAAGAGTAC

[1974] CGTCTGGCGACCAACGCTTCGCAGGCGGGTGTTGAGAAAATTCTGAGCGCGTTGGA

[1975] GATCCCTGATGTCGGTAATCTGAGCCAAGTCGTGGTTATGAAGAGCAAGAACGACC

[1976] AGGGTATCACTAACAAGTGCAAGATGAACCTGCAAGACAACAATGGTAACGACATC

[1977] GGCTTTATTGGTTaCaAaCAGTTCAACAATATTGCTAAACTGGTAGCGAGCAATTGGT

[1978] ACAATCGTCAGATTGAGCGCAGCAGCCGTACTcatGGCTGTAGCTGGGAGTTTATCCC

[1979] GGTCGATGATGGTTGGGGCGAACGTCCGCTGCACCATCACCATCACCAT

[1980] SEQ ID NO:50-rH C The polypeptide sequence (His-tag) of variant Y1117V F1252Y H1253KL1278H.

[1981] MIINTSILNLRYESNHLIDLSRYASKINIGSKVNFDPIDKNQIQLFNLESSKIEVILKNAIVY

[1982] NSMYENFSTSFWIRIPKYFNSISLNNEYTIINCMENNSGWKVSLNYGEIIWTLQDTQEIKQ

[1983] RVVFKYSQMINISDYINRWIFVTITNNRLNNSKIYINGRLIDQKPISNLGNIHASNNIMFKL

[1984] DGCRDTHRYIWIKYFNLFDKELNEKEIKDLYDNQSNSGILKDFWGDYLQYDKPYYMLN

[1985] LVDPNKYVDVNNVGIRGYMYLKGPRGSVMTTNIYLNSSLYRGTKFIIKKYASGNKDNI

[1986] VRNNDRVYINVVVKNKEYRLATNASQAGVEKILSALEIPDVGNLSQVVVMKSKNDQGI

[1987] TNKCKMNLQDNNGNDIGFIGYKQFNNIAKLVASNWYNRQIERSSRTHGCSWEFIPVDD

[1988] GWGERPLHHHHHH

[1989] The polypeptide sequence of SEQ ID NO:51-BoNT / A-UniProtP10845

[1990] MPFVNKQFNYKDPVNGVDIAYIKIPNVGQMQPVKAFKIHNKIWVIPERDTFTNPEEGDL

[1991] NPPPEAKQVPVSYYDSTYLSTDNEKDNYLKGVTKLFERIYSTDLGRMLLTSIVRGIPFW

[1992] GGSTIDTELKVIDTNCINVIQPDGSYRSEELNLVIIGPSADIIQFECKSFGHEVLNLTRNGY

[1993] GSTQYIRFSPDFTFGFEESLEVDTNPLLGAGKFATDPAVTLAHELIHAGHRLYGIAINPNR

[1994] VFKVNTNAYYEMSGLEVSFEELRTFGGHDAKFIDSLQENEFRLYYYNKFKDIASTLNKA

[1995] KSIVGTTASLQYMKNVFKEKYLLSEDTSGKFSVDKLKFDKLYKMLTEIYTEDNFVKFFK

[1996] VLNRKTYLNFDKAVFKINIVPKVNYTIYDGFNLRNTNLAANFNGQNTEINNMNFTKLK

[1997] NFTGLFEFYKLLCVRGIITSKTKSLDKGYNKALNDLCIKVNNWDLFFSPSEDNFTNDLN

[1998] KGEEITSDTNIEAAEENISLDLIQQYYLTFNFDNEPENISIENLSSDIIGQLELMPNIERFPN

[1999] GKKYELDKYTMFHYLRAQEFEHGKSRIALTNSVNEALLNPSRVYTFFSSDYVKKVNKA

[2000] TEAAMFLGWVEQLVYDFTDETSEVSTTDKIADITIIIPYIGPALNIGNMLYKDDFVGALIF

[2001] SGAVILLEFIPEIAIPVLGTFALVSYIANKVLTVQTIDNALSKRNEKWDEVYKYIVTNWL

[2002] AKVNTQIDLIRKKMKEALENQAEATKAIINYQYNQYTEEEKNNINFNIDDLSSKLNESIN

[2003] KAMININKFLNQCSVSYLMNSMIPYGVKRLEDFDASLKDALLKYIYDNRGTLIGQVDRL

[2004] KDKVNNTLSTDIPFQLSKYVDNQRLLSTFTEYIKNIINTSILNLRYESNHLIDLSRYASKIN

[2005] IGSKVNFDPIDKNQIQLFNLESSKIEVILKNAIVYNSMYENFSTSFWIRIPKYFNSISLNNE

[2006] YTIINCMENNSGWKVSLNYGEIIWTLQDTQEIKQRVVFKYSQMINISDYINRWIFVTITN

[2007] NRLNNSKIYINGRLIDQKPISNLGNIHASNNIMFKLDGCRDTHRYIWIKYFNLFDKELNE

[2008] KEIKDLYDNQSNSGILKDFWGDYLQYDKPYYMLNLYDPNKYVDVNNVGIRGYMYLK

[2009] GPRGSVMTTNIYLNSSLYRGTKFIIKKYASGNKDNIVRNNDRVYINVVVKNKEYRLATN

[2010] ASQAGVEKILSALEIPDVGNLSQVVVMKSKNDQGITNKCKMNLQDNNGNDIGFIGFHQ

[2011] FNNIAKLVASNWYNRQIERSSRTLGCSWEFIPVDDGWGERPL

[2012] The polypeptide sequence of SEQ ID NO:52-BoNT / B-UniProtP10844

[2013] MPVTINNFNYNDPIDNNNIIMMEPPFARGTGRYYKAFKITDRIWIIPERYTFGYKPEDFN

[2014] KSSGIFNRDVCEYYDPDYLNTNDKKNIFLQTMIKLFNRIKSKPLGEKLLEMIINGIPYLGD

[2015] RRVPLEEFNTNIASVTVNKLISNPGEVERKKGIFANLIIFGPGPVLNENETIDIGIQNHFAS

[2016] REGFGGIMQMKFCPEYVSVFNNVQENKGASIFNRRGYFSDPALILMHELIHVLHGLYGI

[2017] KVDDLPIVPNEKKFFMQSTDAIQAEELYTFGGQDPSIITPSTDKSIYDKVLQNFRGIVDRL

[2018] NKVLVCISDPNININIYKNKFKDKYKFVEDSEGKYSIDVESFDKLYKSLMFGFTETNIAE

[2019] NYKIKTRASYFSDSLPPVKIKNLLDNEIYTIEEGFNISDKDMEKEYRGQNKAINKQAYEEI

[2020] SKEHLAVYKIQMCKSVKAPGICIDVDNEDLFFIADKNSFSDDLSKNERIEYNTQSNYIEN

[2021] DFPINELILDTDLISKIELPSENTESLTDFNVDVPVYEKQPAIKKIFTDENTIFQYLYSQTFP

[2022] LDIRDISLTSFDDALLFSNKVYSFFSMDYIKTANKVVEAGLFAGWVKQIVNDFVIEANK

[2023] SNTMDKIADISLIVPYIGLALNVGNETAKGNFENAFEIAGASILLEFIPELLIPVVGAFLLE

[2024] SYIDNKNKIIKTIDNALTKRNEKWSDMYGLIVAQWLSTVNTQFYTIKEGMYKALNYQA

[2025] QALEEIIKYRYNIYSEKEKSNINIDFNDINSKLNEGINQAIDNINNFINGCSVSYLMKKMIP

[2026] LAVEKLLDFDNTLKKNLLNYIDENKLYLIGSAEYEKSKVNKYLKTIMPFDLSIYTNDTILI

[2027] EMFNKYNSEILNNIILNLRYKDNNLIDLSGYGAKVEVYDGVELNDKNQFKLTSSANSKI

[2028] RVTQNQNIIFNSVFLDFSVSFWIRIPKYKNDGIQNYIHNEYTIINCMKNNSGWKISIRGNRI

[2029] IWTLIDINGKTKSVFFEYNIREDISEYINRWFFVTITNNLNNAKIYINGKLESNTDIKDIRE

[2030] VIANGEIIFKLDGDIDRTQFIWMKYFSIFNTELSQSNIEERYKIQSYSEYLKDFWGNPLMY

[2031] NKEYYMFNAGNKNSYIKLKKDSPVGEILTRSKYNQNSKYINYRDLYIGEKFIIRRKSNSQ

[2032] SINDDIVRKEDYIYLDFFNLNQEWRVYTYKYFKKEEEKLFLAPISDSDEFYNTIQIKEYDE

[2033] QPTYSCQLLFKKDEESTDEIGLIGIHRFYESGIVFEEYKDYFCISKWYLKEVKRKPYNLKL

[2034] GCNWQFIPKDEGWTE

[2035] The polypeptide sequence of SEQ ID NO:53-BoNT / C-UniProtP18640

[2036] MPITINNFNYSDPVDNKNILYLDTHLNTLANEPEKAFRITGNIWVIPDRFSRNSNPNLNKP

[2037] PRVTSPKSGYYDPNYLSTDSDKDPFLKEIIKLFKRINSREIGEELIYRLSTDIPFPGNNNTPI

[2038] NTFDFDVDFNSVDVKTRQGNNWVKTGSINPSVIITGPRENIIDPETSTFKLTNNTFAAQE

[2039] GGFALSIISISPRFMLTYSNATNDVGEGRFSKSEFCMDPILILMHELNHAMHNLYGIAIPN

[2040] DQTISSVTSNIFYSQYNVKLEYAEIYAFGGPTIDLIPKSARKYFEEKALDYYRSIAKRLNSI

[2041] TTANPSSFNKYIGEYKQKLIRKYRFVVESSGEVTVNRNKFVELYNELTQIFTEFNYAKIY

[2042] NVQNRKIYLSNVYTPVTANILDDNVYDIQNGFNIPKSNLNVLFMGQNLSRNPALRKVNP

[2043] ENMLYLFTKFCHKAIDGRSLYNKTLDCRELLVKNTDLPFIGDISDVKTDIFLRKDINEET

[2044] EVIYYPDNVSVDQVILSKNTSEHGQLDLLYPSIDSESEILPGENQVFYDNRTQNVDYLNS

[2045] YYYLESQKLSDNVEDFFTTRSIEEALDNSAKVYTYFPTLANKVNAGVQGGLFLMWAND

[2046] VVEDFTTNILRKDTLDKISDVSAIIPYIGPALNISNSVRRGNFTEAFAVTGVTILLEAFPEF

[2047] TIPALGAFVIYSKVQERNEIIKTIDNCLEQRIKRWKDSYEWMMGTWLSRIITQFNNISYQ

[2048] MYDSLNYQAGAIKAKIDLEYKKYSGSDKENIKSQVENLKNSLDVKISEAMNNINKFIRE

[2049] CSVTYLFKNMLPKVIDELNEFDRNTKAKLINLIDSHNIILVGEVDKLKAKVNNSFQNTIP

[2050] FNIFSYTNNSLLKDIINEYFNNINDSKILSLQNRKNTLVDTSGYNAEVSEEGDVQLNPIFPF

[2051] DFKLGSSGEDRGKVIVTQNENIVYNSMYESFSISFWIRINKWVSNLPGYTIIDSVKNNSG

[2052] WSIGIISNFLVFTLKQNEDSEQSINFSYDISNNAPGYNKWFFVTVTNNMMGNMKIYINGK

[2053] LIDTIKVKELTGINFSKTITFEINKIPDTGLITSDSDNINMWIRDFYIFAKELDGKDINILFNS

[2054] LQYTNVVKDYWGNDLRYNKEYYMVNIDYLNRYMYANSRQIVFNTRRNNNDFNEGYK

[2055] IIIKRIRGNTNDTRVRGGDILYFDMTINNKAYNLFMKNETMYADNHSTEDIYAIGLREQT

[2056] KDINDNIIFQIQPMNNTYYYASQIFKSNFNGENISGICSIGTYRFRLGGDWYRHNYLVPTV

[2057] KQGNYASLLESTSTHWGFVPVSE

[2058] The polypeptide sequence of SEQ ID NO:54-BoNT / D-UniProtP19321

[2059] MTWPVKDFNYSDPVNDNDILYLRIPQNKLITTPVKAFMITQNIWVIPERFSSDTNPSLSKP

[2060] PRPTSKYQSYYDPSYLSTDEQKDTFLKGIIKLFKRINERDIGKKLINYLVVGSPFMGDSST

[2061] PEDTFDFTRHTTNIAVEKFENGSWKVTNIITPSVLIFGPLPNILDYTASLTLQGQQSNPSFE

[2062] GFGTLSILKVAPEFLLTFSDVTSNQSSAVLGKSIFCMDPVIALMHELTHSLHQLYGINIPS

[2063] DKRIRPQVSEGFFSQDGPNVQFEELYTFGGLDVEIIPQIERSQLREKALGHYKDIAKRLN

[2064] NINKTIPSSWISNIDKYKKIFSEKYNFDKDNTGNFVVNIDKFNSLYSDLTNVMSEVVYSS

[2065] QYNVKNRTHYFSRHYLPVFANILDDNIYTIRDGFNLTNKGFNIENSGQNIERNPALQKLS

[2066] SESVVDLFTKVCLRLTKNSRDDSTCIKVKNNRLPYVADKDSISQEIFENKIITDETNVQN

[2067] YSDKFSLDESILDGQVPINPEIVDPLLPNVNMEPLNLPGEEIVFYDDITKYVDYLNSYYYL

[2068] ESQKLSNNVENITLTTSVEEALGYSNKIYTFLPSLAEKVNKGVQAGLFLNWANEVVEDF

[2069] TTNIMKKDTLDKISDVSVIIPYIGPALNIGNSALRGNFNQAFATAGVAFLLEGFPEFTIPAL

[2070] GVFTFYSSIQEREKIIKTIENCLEQRVKRWKDSYQWMVSNWLSRITTQFNHINYQMYDS

[2071] LSYQADAIKAKIDLEYKKYSGSDKENIKSQVENLKNSLDVKISEAMNNINKFIRECSVTY

[2072] LFKNMLPKVIDELNKFDLRTKTELINLIDSHNIILVGEVDRLKAKVNESFENTMPFNIFSY

[2073] TNNSLLKDIINEYFNSINDSKILSLQNKKNALVDTSGYNAEVRVGDNVQLNTIYTNDFKL

[2074] SSSGDKIIVNLNNNILYSAIYENSSVSFWIKISKDLTNSHNEYTIINSIEQNSGWKLCIRNG

[2075] NIEWILQDVNRKYKSLIFDYSESLSHTGYTNKWFFVTITNNIMGYMKLYINGELKQSQKI

[2076] EDLDEVKLDKTIVFGIDENIDENQMLWIRDFNIFSKELSNEDINIVYEGQILRNVIKDYWG

[2077] NPLKFDTEYYIINDNYIDRYIAPESNVLVLVQYPDRSKLYTGNPITIKSVSDKNPYSRILN

[2078] GDNIILHMLYNSRKYMIIRDTDTIYATQGGECSQNCVYALKLQSNLGNYGIGIFSIKNIVS

[2079] KNKYCSQIFSSFRENTMLLADIYKPWRFSFKNAYTPVAVTNYETKLLSTSSFWKFISRDP

[2080] GWVE

[2081] The polypeptide sequence of SEQ ID NO:55-BoNT / E-UniProtQ00496

[2082] MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTTPQDFHPPTSL

[2083] KNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNLSGGILLEELSKANPYLGNDNTPD

[2084] NQFHIDGADASAVEIKFSNGSQDILLPNVIIMGAEPDLFETNSSNISLRNNYMPSNHRFGSIAI

[2085] VTFSPEYSFRFNDNCMNEFIQDPALTLMHELIHSLHGLYGAKGITTKYTITQKQNPLITNI

[2086] RGTNIEEFLTFGGTDLNIITSAQSNDIYTNLLADYKKIASKLSKVQVSNPLLNPYKDVFEA

[2087] KYGLDKDASGIYSVNINKFNDIFKKLYSFTEFDLRTKFQVKCRQTYIGQYKYFKLSNLL

[2088] NDSIYNISEGYNINNLKVNFRGQNANLNPRIITPITGRGLVKKIIRFCKNIVSVKGIRKSICI

[2089] EINGLEFFVASENSYNDDNINTPKEIDDTVTSNNNYENDLDQVILNFNSESAPGLSDEK

[2090] LNLTIQNDAYIPKYDSNGTSDIEQHDVNELNVFFYLDAQKVPEGENNVNLTSSIDTALLE

[2091] QPKIYTFFSSEFINNVNKPVQAALFVSWIQQVLVDFTTEANQKSTVDKIADISIVVPYIGL

[2092] ALNIGNEAQKGNFKDALELLGAGILLEFEPELLIPTILVFTIKSFLGSSDNKNKVIKAINNA

[2093] LKERDEKWKEVYSFIVSNWMTKINTQFNKRKEQMYQALQNQVNAIKTIIESKYNSYTL

[2094] EEKNELTNKYDIKQIENELNQKVSIAMNNIDRFLTESSISYLMKIINEVKINKLREYDENV

[2095] KTYLLNYIIQHGSILGESQQELNSMVTDTLNNSIPFKLSSYTDDKILISYFNKFFKRIKSSS

[2096] VLNMRYKNDKYVDTSGYDSNININGDVYKYPTNKNQFGIYNDKLSEVNISQNDYIIYDN

[2097] KYKNFSISFWVRIPNYDNKIVNVNNEYTIINCMRDNNSGWKVSLNHNEIIWTFEDNRGI

[2098] NQKLAFNYGNANGISDYINKWIFVTITNDRLGDSKLYINGNLIDQKSILNLGNIHVSDNIL

[2099] FKIVNCSYTRYIGIRIFNIFDKELDETEIQTLYSNEPNTNILKDFWGNYLLYDKEYYLLN

[2100] VLKPNFIDRRKDSTLSINNIRSTILLANNRLYSGIKVKIQRVNNSSTNDNLVRKNDQVYIN

[2101] FVASKTHLFPLYADTATTNKEKTIKISSSSGNRFNQVVVMNSVGNCTMNFKNNNGNNIG

[2102] LLGFKADTVVASTWYYTHMRDHTNSNGCFWNFISEEHGWQEK

[2103] The polypeptide sequence of SEQ ID NO:56-BoNT / F-UniProtA7GBG3

[2104] MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPERNTIGTDPSDFD

[2105] PPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRINSNPAGEVLLQEISYAKPYLGN

[2106] EHTPINEFHPPVTRTTSVNIKSSTVNKSSIILNLLVLGAGPDIHENSSYPVRKLMDSGGVYD

[2107] PSNDGFGSINIVTFSPEYETFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGARGVT

[2108] YKETIKVKQAPLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRV

[2109] NSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFTEIDLANKFKV

[2110] KCRNTYFIKYGFLKVPNLLDDDIYTVSEGFNIGNLAVNNRGQNIKLNPKIIDSIPDKGLVE

[2111] KIVKFCKSVIPRKGTKAPPRLCIRVNNRELFFVASESSYNDINTKEIDDTTTNLNNYR

[2112] NNLDEVILDYNSETIPQISNQTLNTLVQDDSYVPRYDSNGTSEIEEHNVVDLNVFFYLHA

[2113] QKVPEGETNISLTSSIDTALSEESQVYTFFSSEFINTINKPVHAALFISWINQVIRDFTTEAT

[2114] QKSTFDKIADISLVVPYVGLALNIGNEVQKENFKEAFELLGAGILLEFVPELLIPTILVFTI

[2115] KSFIGSSENKNKIIKAINNSLMERETKWKEIYSWIVSNWLTRINTQFNRKKEQMYQALQ

[2116] NQVDAIKTVIEYKYNNYTSDERNRLESEYNINNIREELNKKVSLAMENIERFITESSIFYL

[2117] MKLINEAKVSKLREYDEGVKEYLLDYISEHRSILGNSVQELNDLVTSTLNNSIPFELSSYT

[2118] NDKILILYFNKLYKKIKDNSILDMRYENNKFIDISGYGSNISINGDVYIYSTNRNQFGIYSS

[2119] KPSEVNIAQNNDIIYNGRYQNFSISFWVRIPKYFNKVNLNNEYTIIDCIRNNNSGWKISLN

[2120] YNKIIWTLQDTAGNNQKLVFNYTQMISISDYINKWIFVTITNNRLGNSRIYINGNLIDEKS

[2121] ISNLGDIHVSDNILFKIVGCNDTRYVGIRYFKVFDTELGKTEIETLYSDEPDPSILKDFWG

[2122] NYLLYNKRYYLLNLLRTDKSITQNSNFLNINQQRGVYQKPNIFSNTRLYTGVEVIIRKNG

[2123] STDISNTDNFVRKNDLAYINVVDRDVEYRLYADISIAKPEKIIKLIRTSNSNNSLGQIIVM

[2124] DSIGNNCTMNFQNNNGGNIGLLGFHSNNLVASSWYYNNIRKNTSSNGCFWSFISKEHG

[2125] WQEN

[2126] The polypeptide sequence of SEQ ID NO:57-BoNT / G-UniProtQ60393

[2127] MPVNIKXFNYNDPINNDDIIMMEPFNDPGPGTYYKAFRIIDRIWIVPERFTYGFQPDQFN

[2128] ASTGVFSKDVYEYYDPTYLKTDAEKDKFLKTMIKLFNRINSKPSGQRLLDMIVDAIPYL

[2129] GNASTPPDKFAANVANVSINKKIIQPGAEDQIKGLMTNLIIFGPGPVLSDNFTDSMIMNG

[2130] HSPISEGFGARMMIRFCPSCLNVFNNVKENDTSIFSRRAYFADPALTLMHELIHVLHGL

[2131] YGIKISNLPITPNTKEFFMQHSDPVQAEELYTFGGHDPSVISPSTDMNIYNKALQNFQDIA

[2132] NRLNIVSSAQGSGIDISLYKQIYKNKYDFVEDPNGKYSVDKDKFDKLYKALMFGFTETN

[2133] LAGEYGIKTRYSYFSYLPPIKTEKLLDNTIYTQNEGFNIASKNLKTEFNGQNKAVNKEA

[2134] YEEISLEHLVIYRIAMCKPVMYKNTGKSEQCIIVNNEDLFFIANKDSFSKDLAKAETIAY

[2135] NTQNNTIENNFSIDQLILDNDLSSGIDLPNETPFTNFDDIDIPVYIKQSALKKIFVDGDS

[2136] LFEYLHAQTFPSNIENLQLTNSLNDALRNNNKVYTFFSTNLVEKANTVVGASLFVNWV

[2137] KGVIDDFTSESTQKSTIDKVSDVSIIIPYIGPALNVGNETAKENFKNAFEIGGAAILMEFIP

[2138] ELIVPIVGFFTLESYVGNKGHIIMTISNALKKRDQKWTDMYGLIVSQWLSTVNTQFYTIK

[2139] ERMYNALNNQSQAIEKIIEDQYNRYSEEDKMNINIDFNDIDFKLNQSINLAINNIDDFINQ

[2140] CSISYLMNRMIPLAVKKLKDFDDNLKRDLLEYIDTNELYLLDEVNILKSKVNRHLKDSIP

[2141] FDLSLYTKDTILIQVFNNYISNISSNAILSSYRGGRLIDSSGYGATMNVGSDVIFNDIGNG

[2142] QFKLNNSENSNITAHQSKFVVYDSMFDNFSINFWVRTPKYNNDIQTYLQNEYTIISCIK

[2143] NDSGWKVSIKGNRIIWTLIDVNAKSKSIFFEYSIKDNISDYINKWFSITITNDRLGNANIYI

[2144] NGSLKKSEKILNLDRINSSSNDIDFKLINCTDTTKFVWIKDFNIFGRELNATEVSSLYWIQS

[2145] STNTLKDFWGNPLRYDTQYYLFNQGMQNIYIKIFSKASMGETAPRTNFNNAAINYQNL

[2146] YLGLRFIIKKASNSRNINNDNIVREGDYIYLNIDNISDESYRVYVLVNSKEIQTQLFLAPIN

[2147] DDPTFYDVLQIKKYYEKTTYNCQILCEKDTKTFGLFGIGKFWKDYGYVWDTYDNYFCIS

[2148] QWYLRRISENINKLRLGCNWQFIPVDEGWTE

[2149] The polypeptide sequence of SEQ ID NO:58-TeNT-UniProtP04958

[2150] MPITINNFRYSDPVNNDTIIMMEPPYCKGLDIYYKAFKITTRIWIVPERYEFGTKPEDFNP

[2151] PSSLIEGASEYYDPNYLRTDSDKDRFLQTMVKLFNRIKNNVAGEALLDKIINAIPYLGNS

[2152] YSLLDKFDTNSNSVSFNLLEQDPSGATTKSAMLTNLIIFGPGPVLNKNEVRGIVLRVDNK

[2153] NYFPCRDGFGSIMQMAFCPEYVPTFDNVIENITSLTIGKSKYFQDPALLLMHELIHVLHG

[2154] LYGMQVSSHEIIPSKQEIYMQHTYPISAEELFTFGGQDANLISIDIKNDLYEKTLNDYKAI

[2155] ANKLSQVTSCNDPNIDIDSYKQIYQQKYQFDKDSNGQYIVNEDKFQILYNSIMYGFTEIE

[2156] LGKKFNIKTRLSYFSMNHDPVKIPNLLDDTIYNDTEGFNIESKDLKSEYKGQNMRVNTN

[2157] AFRNVDGSGLVSKLIGLCKKIIPPTNIRENLYNRTASLTDLGGELCIKIKNEDLTFIAEKNS

[2158] FSEEPFQDEIVSYNTKNKPLNFNYSLDKIIVDYNLQSKITLPNDRTTPVTKGIPYAPEYKS

[2159] NAASTIEIHNIDDNTIYQYLYAQKSPTTLQRITMTNSVDDALINSTKIYSYFPSVISKVNQ

[2160] GAQGILFLQWVRDIIDDFTNESSQKTTIDKISDVSTIVPYIGPALNIVKQGYEGNFIGALET

[2161] TGVVLLLEYIPEITLPVIAALSIAESSTQKEKIIKTIDNFLEKRYEKWIEVYKLVKAKWLGT

[2162] VNTQFQKRSYQMYRSLEYQVDAIKKIIDYEYKIYSGPDKEQIADEINNLKNKLEEKANK

[2163] AMININIFMRESSRSFLVNQMINEAKKQLLEFDTQSKNILMQYIKANSKFIGITELKKLES

[2164] KINKVFSTPIPFSYSKNLDCWVDNEEDIDVILKKSTILNLDINNDIISDISGFNSSVITYPDA

[2165] QLVPGINGKAIHLVNNESSEVIVHKAMDIEYNDMFNNFTVSFWLRVPKVSASHLEQYGT

[2166] NEYSIISSMKKHSLSIGSGWSVSLKGNNLIWTLKDSAGEVRQITFRDLPDKFNAYLANK

[2167] WVFITITNDRLSSANLYINGVLMGSAEITGLGAIREDNNITLKLDRCNNNNQYVSIDKFRI

[2168] FCKALNPKEIEKLYTSYLSITFLRDFWGNPLRYDTEYYLIPVASSSKDVQLKNIDTYMYL

[2169] TNAPSYTNGKLNIYYRRLYNGLKFIIKRYTPNNEIDSFVKSGDFIKLYVSYNNNEHIVGY

[2170] PKDGNAFNNLDRILRVGYNAPGIPLYKKMEAVKLRDLKTYSVQLKLYDDKNASLGLV

[2171] GTHNGQIGNDPNRDILIASNWYFNHLKDKILGCDWYFVPTDEGWTND

[2172] The polypeptide sequence of SEQ IDNO:59-BoNT / X

[2173] MKLEINKFNYNDPIDGINVITMRPPRHSDKINKGKGPFKAFQVIKNIWIVPERYNFTNNT

[2174] NDLNIPSEPIMEADAIYNPNYLNTPSEKDEFLQGVIKVLERIKSKPEKEKLELLISSSIPL

[2175] VSNGALTLSDNETIAYQENNIVSNLQANLVIYGPGPDIANNATYGLYSTPISNGEGTLS

[2176] EVSFSPFYLKPFDESYGNYRSLVNIVNKFVKREFAPDPASTLMHELVHVTHNLYGISNR

[2177] NFYYNFDTGKIETSRQQNSLIFEELLTFGGIDSKAISSLIIKKIIETAKNYTLLISERLNTV

[2178] TVENDLLKYIKNKIPVQGRLGNFKLDTAEFEKKLNTILFVLNESNLAQRFSILVRKHYLK

[2179] ERPIDPIYVNILDDNSYSTLEGFNISSQGSNDFQGQLLLESSYFEKIESNALRAFIKICPRNGL

[2180] LYNAIYRNSKNYLNNIDLEDKKTTSKTNVSYPCSLLNGCIEVENKDLFLISNKDSLNDIN

[2181] LSEEKIKPETTVFFKDKLPPQDITLSNYDFTEANSIPSISQQNILERNEELYEPIRNSLFEIKT

[2182] IYVDKLTTFHFLEAQNIDESIDESKIRVELTDSVDEALSNPNKVYSPFKNMNSTINSIETGI

[2183] TSTYIFYQWLRSIVKDFSDETGKIDVIDKSSDTLAIVPYIGPLLNIGNDIRHGDFVGAIELA

[2184] GITALLEYVPEFTIPILVGLEVIGGELAREQVEAIVNNALDKRDQKWAEVYNITKAQWW

[2185] GTIHLQINTRLAHTYKALSRQANAIKMNMEFQLANYKGNIDDKAKIKNAISETEILNKS

[2186] VEGAMKNTEKFMIKLSNSYLTKEMIPKVQDNLKNFDLETKKTLDKFIKEKEDILGTNLS

[2187] SSLRRKVSIRLNKNIAFDINDIPFSEFDDLINQYKNEIEDYEVLNLGAEDGKIKDLSGTTSD

[2188] INIGSDIELADGRENKAIKIKGSENSTIKIAMNKYLRFSATDNFSISFWIKHPKPTNLLNNG

[2189] IEYTLVENFNQRGWKISIQDSKLIWYLRDHNNSIKIVTPDYIAFNGWNLITITNNRSKGSI

[2190] VYVNGSKIEEKDISSIWNTEVDDPIIFRLKNNRDTQAFTLLDQFSIYRKELNQNEVVKLY

[2191] NYYFNSNYIRDIWGNPLQYNKKYYLQTQDKPGKGLIREYWSSFGYDYVILSDSKTITFP

[2192] NNIRYGALYNGSKVLIKNSKKLDGLVRNKDFIQLEIDGYNMGISADRFNEDTNYIGTTY

[2193] GTTHDLTTDFEIIQRQEKYRNYCQLKTPYNIFHKSGLMSTETSKPTFHDYRDWVYSSAW

[2194] YFQNYENLNLRKHTKTNWYFIPKDEGWDED

[2195] The nucleotide sequence of SEQ ID NO:60-mrBoNT / A

[2196] ATGCCATTCGTCAACAAGCAATTCAACTACAAAGACCCAGTCAACGGCGTCGACAT

[2197] CGCATACATCAAGATTCCGAACGCCGGTCAAATGCAGCCGGTTAAGGCTTTTAAGA

[2198] TCCACAACAAGATTTGGGTTATCCCGGAGCGTGACACCTTCACGAACCCGGAAGAA

[2199] GGCGATCTGAACCCGCCACCGGAAGCGAAGCAAGTCCCTGTCAGCTACTACGATTC

[2200] GACGTACCTGAGCACGGATAACGAAAAAGATAACTACCTGAAAGGTGTGACCAAG

[2201] CTGTTCGAACGTATCTACAGCACGGATCTGGGTCGCATGCTGCTGACTAGCATTGTT

[2202] CGCGGTATCCCGTTCTGGGGTGGTAGCACGATTGACACCGAACTGAAGGTTATCGA

[2203] CACTAACTGCATTAACGTTATTCAACCGGATGGTAGCTATCGTAGCGAAGAGCTGA

[2204] ATCTGGTCATCATTGGCCCGAGCGCAGACATTATCCAATTCGAGTGCAAGAGCTTTG

[2205] GTCACGAGGTTCTGAATCTGACCCGCAATGGCTATGGTAGCACCCAGTACATTCGTT

[2206] TTTCGCCGGATTTTACCTTCGGCTTTGAAGAGAGCCTGGAGGTTGATACCAATCCGT

[2207] TGCTGGGTGCGGGCAAATTCGCTACCGATCCGGCTGTCACGCTGGCCCATGAACTG

[2208] ATCCACGCAGGCCACCGCCTGTACGGCATTGCCATCAACCCAAACCGTGTGTTCAA

[2209] GGTTAATACGAATGCATACTACGAGATGAGCGGCCTGGAAGTCAGCTTCGAAGAAC

[2210] TGCGCACCTTCGGTGGCCATGACGCTAAATTCATTGACAGCTTGCAAGAGAATGAG

[2211] TTCCGTCTGTACTACTATAACAAATTCAAAGACATTGCAAGCACGTTGAACAAGGC

[2212] CAAAAGCATCGTTGGTACTACCGCGTCGTTGCAGTATATGAAGAATGTGTTTAAAG

[2213] AGAAGTACCTGCTGTCCGAGGATACCTCCGGCAAGTTTAGCGTTGATAAGCTGAAG

[2214] TTTGACAAACTGTACAAGATGCTGACCGAGATTTACACCGAGGACAACTTTGTGAA

[2215] ATTCTTCAAAGTGTTGAATCGTAAAACCTATCTGAATTTTGACAAAGCGGTTTTCAA

[2216] GATTAACATCGTGCCGAAGGTGAACTACACCATCTATGACGGTTTTAACCTGCGTAA

[2217] CACCAACCTGGCGGCGAACTTTAACGGTCAGAATACGGAAATCAACAACATGAATT

[2218] TCACGAAGTTGAAGAACTTCACGGGTCTGTTCGAGTTCTATAAGCTGCTGTGCGTGC

[2219] GCGGTATCATCACCAGCAAAACCAAAAGCCTGGACAAAGGCTACAACAAGGCGCT

[2220] GAATGACCTGTGCATTAAGGTAAACAATTGGGATCTGTTCTTTTCGCCATCCGAAGA

[2221] TAATTTTACCAACGACCTGAACAAGGGTGAAGAAATCACCAGCGATACGAATATTG

[2222] AAGCAGCGGAAGAGAATATCAGCCTGGATCTGATCCAGCAGTACTATCTGACCTTT

[2223] AACTTCGACAATGAACCGGAGAACATTAGCATTGAGAATCTGAGCAGCGACATTAT

[2224] CGGTCAGCTGGAACTGATGCCGAATATCGAACGTTTCCCGAACGGCAAAAAGTACG

[2225] AGCTGGACAAGTACACTATGTTCCATTACCTGCGTGCACAGGAGTTTGAACACGGT

[2226] AAAAGCCGTATCGCGCTGACCAACAGCGTTAACGAGGCCCTGCTGAACCCGAGCCG

[2227] TGTCTATACCTTCTTCAGCAGCGACTATGTTAAGAAAGTGAACAAAGCCACTGAGG

[2228] CCGCGATGTTCCTGGGCTGGGTGGAACAGCTGGTATATGACTTCACGGACGAGACG

[2229] AGCGAAGTGAGCACTACCGACAAAATTGCTGATATTACCATCATTATCCCGTATATT

[2230] GGTCCGGCACTGAACATTGGCAACATGCTGTACAAAGACGATTTTGTGGGTGCCCT

[2231] GATCTTCTCCGGTGCCGTGATTCTGCTGGAGTTCATTCCGGAGATTGCGATCCCGGT

[2232] GTTGGGTACCTTCGCGCTGGTGTCCTACATCGCGAATAAGGTTCTGACGGTTCAGAC

[2233] CATCGATAACGCGCTGTCGAAACGTAATGAAAAATGGGACGAGGTTTACAAATACA

[2234] TTGTTACGAATTGGCTGGCGAAAGTCAATACCCAGATCGACCTGATCCGTAAGAAA

[2235] ATGAAAGAGGCGCTGGAGAATCAGGCGGAGGCCACCAAAGCAATTATCAACTACC

[2236] AATACAACCAGTACACGGAAGAAGAGAAGAATAACATTAACTTCAATATCGATGAT

[2237] TTGAGCAGCAAGCTGAATGAATCTATCAACAAAGCGATGATCAATATCAACAAGTT

[2238] TTTGAATCAGTGTAGCGTTTCGTACCTGATGAATAGCATGATTCCGTATGGCGTCAA

[2239] ACGTCTGGAGGACTTCGACGCCAGCCTGAAAGATGCGTTGCTGAAATACATTTACG

[2240] ACAATCGTGGTACGCTGATTGGCCAAGTTGACCGCTTGAAAGACAAAGTTAACAAT

[2241] ACCCTGAGCACCGACATCCCATTTCAACTGAGCAAGTATGTTGATAATCAACGTCTG

[2242] TTGAGCACTTTCACCGAGTATATCAAAAACATCATCAATACTAGCATTCTGAACCTG

[2243] CGTTACGAGAGCAAGCATCTGATTGATCTGAGCCGTTATGCTAGCAAGATCAACAT

[2244] CGGTAGCAAGGTCAATTTTGACCCGATCGATAAGAACCAGATCCAGCTGTTTAATCT

[2245] GGAATCGAGCAAAATTGAGGTTATCCTGAAAAAGGCCATTGTCTACAACTCCATGT

[2246] ACGAGAATTTCTCCACCAGCTTCTGGATTCGCATCCCGAAATACTTCAACAAGATTA

[2247] GCCTGAACAACGAGTATACTATCATCAACTGTATGGAGAACAACAGCGGTTGGAAG

[2248] GTGTCTCTGAACTATGGTGAGATCATTTGGACCTTGCAGGACACCAAAGAGATCAA

[2249] GCAGCGCGTCGTGTTCAAGTACTCTCAAATGATCAACATTTCCGATTACATTAATCG

[2250] TTGGATCTTCGTGACCATTACGAATAACCGTCTGAATAAGAGCAAGATTTACATCAA

[2251] TGGTCGCTTGATCGATCAGAAACCGATTAGCAACCTGGGTAATATCCACGCAAGCA

[2252] ACAAGATTATGTTCAAATTGGACGGTTGCCGCGATACCCATCGTTATATCTGGATCA

[2253] AGTATTTCAACCTGTTTGATAAAGAACTGAATGAGAAGGAGATCAAAGATTTGTAT

[2254] GACAACCAATCTAACAGCGGCATTTTGAAGGACTTCTGGGGCGATTATCTGCAATA

[2255] CGATAAGCCGTACTATATGCTGAACCTGTATGATCCGAACAAATATGTGGATGTCA

[2256] ATAATGTGGGTATTCGTGGTTACATGTATTTGAAGGGTCCGCGTGGCAGCGTTATGA

[2257] CGACCAACATTTACCTGAACTCTAGCCTGTACCGTGGTACGAAATTCATCATTAAGA

[2258] AATATGCCAGCGGCAACAAAGATAACATTGTGCGTAATAACGATCGTGTCTACATC

[2259] AACGTGGTCGTGAAGAATAAAGAGTACCGTCTGGCGACCAACGCTTCGCAGGCGGG

[2260] TGTTGAGAAAATTCTGAGCGCGTTGGAGATCCCTGATGTCGGTAATCTGAGCCAAG

[2261] TCGTGGTTATGAAGAGCAAGAACGACAAGGGTATCACTAACAAGTGCAAGATGAA

[2262] CCTGCAAGACAACAATGGTAACGACATCGGCTTTATTGGTTTCCACCAGTTCAACAA

[2263] TATTGCTAAACTGGTAGCGAGCAATTGGTACAATCGTCAGATTGAGCGCAGCAGCc

[2264] GTACTTTGGGCTGTAGCTGGGAGTTTATCCCGGTCGATGATGGTTGGGGCGAACGTC

[2265] CGCTG

[2266] The polypeptide sequence of SEQ ID NO:61-mrBoNT / A

[2267] MPFVNKQFNYKDPVNGVDIAYIKIPNAGQMQPVKAFKIHNKIWVIPERDTFTNPEEGDL

[2268] NPPPEAKQVPVSYYDSTYLSTDNEKDNYLKGVTKLFERIYSTDLGRMLLTSIVRGIPFW

[2269] GGSTIDTELKVIDTNCINVIQPDGSYRSEELNLVIIGPSADIIQFECKSFGHEVLNLTRNGY

[2270] GSTQYIRFSPDFTFGFEESLEVDTNPLLGAGKFATDPAVTLAHELIHAGHRLYGIAINPNR

[2271] VFKVNTNAYYEMSGLEVSFEELRTFGGHDAKFIDSLQENEFRLYYYNKFKDIASTLNKA

[2272] KSIVGTTASLQYMKNVFKEKYLLSEDTSGKFSVDKLKFDKLYKMLTEIYTEDNFVKFFK

[2273] VLNRKTYLNFDKAVFKINIVPKVNYTIYDGFNLRNTNLAANFNGQNTEINNMNFTKLK

[2274] NFTGLFEFYKLLCVRGIITSKTKSLDKGYNKALNDLCIKVNNWDLFFSPSEDNFTNDLN

[2275] KGEEITSDTNIEAAEENISLDLIQQYYLTFNFDNEPENISIENLSSDIIGQLELMPNIERFPN

[2276] GKKYELDKYTMFHYLRAQEFEHGKSRIALTNSVNEALLNPSRVYTFFSSDYVKKVNKA

[2277] TEAAMFLGWVEQLVYDFTDETSEVSTTDKIADITIIIPYIGPALNIGNMLYKDDFVGALIF

[2278] SGAVILLEFIPEIAIPVLGTFALVSYIANKVLTVQTIDNALSKRNEKWDEVYKYIVTNWL

[2279] AKVNTQIDLIRKKMKEALENQAEATKAIINYQYNQYTEEEKNNINFNIDDLSSKLNESIN

[2280] KAMININKFLNQCSVSYLMNSMIPYGVKRLEDFDASLKDALLKYIYDNRGTLIGQVDRL

[2281] KDKVNNTLSTDIPFQLSKYVDNQRLLSTFTEYIKNIINTSILNLRYESKHLIDLSRYASKIN

[2282] IGSKVNFDPIDKNQIQLFNLESSKIEVILKKAIVYNSMYENFSTSFWIRIPKYFNKISLNNE

[2283] YTIINCMENNSGWKVSLNYGEIIWTLQDTKEIKQRVVFKYSQMINISDYINRWIFVTITN

[2284] NRLNKSKIYINGRLIDQKPISNLGNIHASNKIMFKLDGCRDTHRYIWIKYFNLFDKELNE

[2285] KEIKDLYDNQSNSGILKDFWGDYLQYDKPYYMLNLYDPNKYVDVNNVGIRGYMYLK

[2286] GPRGSVMTTNIYLNSSLYRGTKFIIKKYASGNKDNIVRNNDRVYINVVVKNKEYRLATN

[2287] ASQAGVEKILSALEIPDVGNLSQVVVMKSKNDKGITNKCKMNLQDNNGNDIGFIGFHQ

[2288] FNNIAKLVASNWYNRQIERSSRTLGCSWEFIPVDDGWGERPL

[2289] SEQ ID NO:62 - Unmodified BoNT / A1 polypeptide sequence

[2290] MPFVNKQFNYKDPVNGVDIAYIKIPNAGQMQPVKAFKIHNKIWVIPERDTFTNPEEGDL

[2291] NPPPEAKQVPVSYYDSTYLSTDNEKDNYLKGVTKLFERIYSTDLGRMLLTSIVRGIPFW

[2292] GGSTIDTELKVIDTNCINVIQPDGSYRSEELNLVIIGPSADIIQFECKSFGHEVLNLTRNGY

[2293] GSTQYIRFSPDFTFGFEESLEVDTNPLLGAGKFATDPAVTLAHELIHAGHRLYGIAINPNR

[2294] VFKVNTNAYYEMSGLEVSFEELRTFGGHDAKFIDSLQENEFRLYYYNKFKDIASTLNKA

[2295] KSIVGTTASLQYMKNVFKEKYLLSEDTSGKFSVDKLKFDKLYKMLTEIYTEDNFVKFFK

[2296] VLNRKTYLNFDKAVFKINIVPKVNYTIYDGFNLRNTNLAANFNGQNTEINNMNFTKLK

[2297] NFTGLFEFYKLLCVRGIITSKTKSLDKGYNKALNDLCIKVNNWDLFFSPSEDNFTNDLN

[2298] KGEEITSDTNIEAAEENISLDLIQQYYLTFNFDNEPENISIENLSSDIIGQLELMPNIERFPN

[2299] GKKYELDKYTMFHYLRAQEFEHGKSRIALTNSVNEALLNPSRVYTFFSSDYVKKVNKA

[2300] TEAAMFLGWVEQLVYDFTDETSEVSTTDKIADITIIIPYIGPALNIGNMLYKDDFVGALIF

[2301] SGAVILLEFIPEIAIPVLGTFALVSYIANKVLTVQTIDNALSKRNEKWDEVYKYIVTNWL

[2302] AKVNTQIDLIRKKMKEALENQAEATKAIINYQYNQYTEEEKNNINFNIDDLSSKLNESIN

[2303] KAMININKFLNQCSVSYLMNSMIPYGVKRLEDFDASLKDALLKYIYDNRGTLIGQVDRL

[2304] KDKVNNTLSTDIPFQLSKYVDNQRLLSTFTEYIKNIINTSILNLRYESNHLIDLSRYASKIN

[2305] IGSKVNFDPIDKNQIQLFNLESSKIEVILKNAIVYNSMYENFSTSFWIRIPKYFNSISLNNE

[2306] YTIINCMENNSGWKVSLNYGEIIWTLQDTQEIKQRVVFKYSQMINISDYINRWIFVTITN

[2307] NRLNNSKIYINGRLIDQKPISNLGNIHASNNIMFKLDGCRDTHRYIWIKYFNLFDKELNE

[2308] KEIKDLYDNQSNSGILKDFWGDYLQYDKPYYMLNLYDPNKYVDVNNVGIRGYMYLK

[2309] GPRGSVMTTNIYLNSSLYRGTKFIIKKYASGNKDNIVRNNDRVYINVVVKNKEYRLATN

[2310] ASQAGVEKILSALEIPDVGNLSQVVVMKSKNDQGITNKCKMNLQDNNGNDIGFIGFHQ

[2311] FNNIAKLVASNWYNRQIERSSRTLGCSWEFIPVDDGWGERPL

[2312] The polypeptide sequence of SEQ ID NO: 63 -mrBoNT / AB

[2313] MPFVNKQFNYKDPVNGVDIAYIKIPNAGQMQPVKAFKIHNKIWVIPERDTFTNPEEGDL

[2314] NPPPEAKQVPVSYYDSTYLSTDNEKDNYLKGVTKLFERIYSTDLGRMLLTSIVRGIPFW

[2315] GGSTIDTELKVIDTNCINVIQPDGSYRSEELNLVIIGPSADIIQFECKSFGHEVLNLTRNGY

[2316] GSTQYIRFSPDFTFGFEESLEVDTNPLLGAGKFATDPAVTLAHELIHAGHRLYGIAINPNR

[2317] VFKVNTNAYYEMSGLEVSFEELRTFGGHDAKFIDSLQENEFRLYYYNKFKDIASTLNKA

[2318] KSIVGTTASLQYMKNVFKEKYLLSEDTSGKFSVDKLKFDKLYKMLTEIYTEDNFVKFFK

[2319] VLNRKTYLNFDKAVFKINIVPKVNYTIYDGFNLRNTNLAANFNGQNTEINNMNFTKLK

[2320] NFTGLFEFYKLLCVRGIITSKTKSLDKGYNKALNDLCIKVNNWDLFFSPSEDNFTNDLN

[2321] KGEEITSDTNIEAAEENISLDLIQQYYLTFNFDNEPENISIENLSSDIIGQLELMPNIERFPN

[2322] GKKYELDKYTMFHYLRAQEFEHGKSRIALTNSVNEALLNPSRVYTFFSSDYVKKVNKA

[2323] TEAAMFLGWVEQLVYDFTDETSEVSTTDKIADITIIIPYIGPALNIGNMLYKDDFVGALIF

[2324] SGAVILLEFIPEIAIPVLGTFALVSYIANKVLTVQTIDNALSKRNEKWDEVYKYIVTNWL

[2325] AKVNTQIDLIRKKMKEALENQAEATKAIINYQYNQYTEEEKNNINFNIDDLSSKLNESIN

[2326] KAMININKFLNQCSVSYLMNSMIPYGVKRLEDFDASLKDALLKYIYDNRGTLIGQVDRL

[2327] KDKVNNTLSTDIPFQLSKYVDNQRLLSTFTEYIKNILNNIILNLRYKDNNLIDLSGYGAK

[2328] VEVYDGVELNDKNQFKLTSSANSKIRVTQNQNIIFNSVFLDFSVSFWIRIPKYKNDGIQN

[2329] YIHNEYTIINCMKNNSGWKISIRGNRIIWTLIDINGKTKSVFFEYNIREDISEYINRWFFVTI

[2330] TNNLNNAKIYINGKLESNTDIKDIREVIANGEIIFKLDGDIDRTQFIWMKYFSIFNTELSQS

[2331] NIEERYKIQSYSEYLKDFWGNPLMYNKEYYMFNAGNKNSYIKLKKDSPVGEILTRSKY

[2332] NQNSKYINYRDLYIGEKFIIRRKSNSQSINDDIVRKEDYIYLDFFNLNQEWRVYTYKYFK

[2333] KEEMKLFLAPIYDSDEFYNTIQIKEYDEQPTYSCQLLFKKDEESTDEIGLIGIHRFYESGIV

[2334] FEEYKDYFCISKWYLKEVKRKPYNLKLGCNWQFIPKDEGWTE

[2335] The polypeptide sequence of SEQ ID NO:64-mrBoNT / AB(0)

[2336] MPFVNKQFNYKDPVNGVDIAYIKIPNAGQMQPVKAFKIHNKIWVIPERDTFTNPEEGDL

[2337] NPPPEAKQVPVSYYDSTYLSTDNEKDNYLKGVTKLFERIYSTDLGRMLLTSIVRGIPFW

[2338] GGSTIDTELKVIDTNCINVIQPDGSYRSEELNLVIIGPSADIIQFECKSFGHEVLNLTRNGY

[2339] GSTQYIRFSPDFTFGFEESLEVDTNPLLGAGKFATDPAVTLAHQLIYAGHRLYGIAINPNR

[2340] VFKVNTNAYYEMSGLEVSFEELRTFGGHDAKFIDSLQENEFRLYYYNKFKDIASTLNKA

[2341] KSIVGTTASLQYMKNVFKEKYLLSEDTSGKFSVDKLKFDKLYKMLTEIYTEDNFVKFFK

[2342] VLNRKTYLNFDKAVFKINIVPKVNYTIYDGFNLRNTNLAANFNGQNTEINNMNFTKLK

[2343] NFTGLFEFYKLLCVRGIITSKTKSLDKGYNKALNDLCIKVNNWDLFFSPSEDNFTNDLN

[2344] KGEEITSDTNIEAAEENISLDLIQQYYLTFNFDNEPENISIENLSSDIIGQLELMPNIERFPN

[2345] GKKYELDKYTMFHYLRAQEFEHGKSRIALTNSVNEALLNPSRVYTFFSSDYVKKVNKA

[2346] TEAAMFLGWVEQLVYDFTDETSEVSTTDKIADITIIIPYIGPALNIGNMLYKDDFVGALIF

[2347] SGAVILLEFIPEIAIPVLGTFALVSYIANKVLTVQTIDNALSKRNEKWDEVYKYIVTNWL

[2348] AKVNTQIDLIRKKMKEALENQAEATKAIINYQYNQYTEEEKNNINFNIDDLSSKLNESIN

[2349] KAMININKFLNQCSVSYLMNSMIPYGVKRLEDFDASLKDALLKYIYDNRGTLIGQVDRL

[2350] KDKVNNTLSTDIPFQLSKYVDNQRLLSTFTEYIKNILNNIILNLRYKDNNLIDLSGYGAK [235...

Claims

1. A polypeptide for promoting neuronal growth or neuronal repair to treat a neurological disorder in a subject, wherein the polypeptide comprises: Clostridial neurotoxin light chain (L-chain) or fragments thereof; and / or Fragments of the heavy chain (H-chain) of Clostridial neurotoxin.

2. A method for promoting neuronal growth or neuronal repair to treat a neurological disorder in a subject, the method comprising administering to the subject a polypeptide, wherein the polypeptide comprises: Clostridial neurotoxin L-chain or fragments thereof; and / or Fragments of the H-chain of Clostridial neurotoxins.

3. Use of a polypeptide in the preparation of a medicament for promoting neuronal growth or neuronal repair to treat a neurological disorder in a subject, wherein the polypeptide comprises: Clostridial neurotoxin L-chain or fragments thereof; and / or Fragments of the H-chain of Clostridial neurotoxins.

4. A polypeptide for use in promoting neuronal growth or neuronal repair to treat a neurological disorder in a subject, wherein the polypeptide comprises a catalytically inactive Clostridial neurotoxin L-chain.

5. A method for promoting neuronal growth or neuronal repair to treat a neurological disorder in a subject, the method comprising administering to the subject a polypeptide, wherein the polypeptide comprises a catalytically inactive Clostridial neurotoxin L-chain.

6. Use of a polypeptide comprising a catalytically inactive Clostridial neurotoxin L-chain in the preparation of a medicament for promoting neuronal growth or neuronal repair to treat a neurological disorder in a subject.

7. A polypeptide for promoting neuronal growth or neuronal repair to treat a neurological disorder in a subject, wherein the polypeptide comprises a polypeptide sequence having at least 70% sequence identity with SEQ ID NO:42 and / or wherein the polypeptide comprises a polypeptide sequence encoded by a nucleotide sequence having at least 70% sequence identity with SEQ ID NO:

41.

8. A method for promoting neuronal growth or neuronal repair to treat a neurological disorder in a subject, the method comprising administering a polypeptide to the subject, wherein the polypeptide comprises a polypeptide sequence having at least 70% sequence identity to SEQ ID NO:42 and / or wherein the polypeptide comprises a polypeptide sequence encoded by a nucleotide sequence having at least 70% sequence identity to SEQ ID NO:

41.

9. Use of a polypeptide in the preparation of a medicament for promoting neuronal growth or neuronal repair to treat a neurological disorder in a subject, wherein the polypeptide comprises a polypeptide sequence having at least 70% sequence identity with SEQ ID NO:42 and / or wherein the polypeptide comprises a polypeptide sequence encoded by a nucleotide sequence having at least 70% sequence identity with SEQ ID NO:

41.

10. A polypeptide for promoting neuronal growth or neuronal repair to treat a neurological disorder in a subject, wherein the polypeptide comprises a polypeptide sequence having at least 70% sequence identity to SEQ ID NO: 63 or 64.

11. A method for promoting neuronal growth or neuronal repair to treat a neurological disorder in a subject, the method comprising administering to the subject a polypeptide, wherein the polypeptide comprises a polypeptide sequence having at least 70% sequence identity to SEQ ID NO: 63 or 64.

12. Use of a polypeptide in the preparation of a medicament for promoting neuronal growth or neuronal repair to treat a neurological disorder in a subject, wherein the polypeptide comprises a polypeptide sequence having at least 70% sequence identity with SEQ ID NO: 63 or 64.

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