Methods and compositions for treating inflammatory skin disease with recombinant microorganisms

Abstract

To provide isolated plasmids, recombinant microorganisms, kits, and methods for the treatment of inflammatory skin disease.SOLUTION: The present invention provides a recombinant microorganism capable of secreting a polypeptide, wherein the recombinant microorganism comprises an expression vector comprising a first coding sequence comprising a gene capable of expressing the polypeptide and a second coding sequence comprising a gene capable of expressing a cell penetrating peptide.SELECTED DRAWING: Figure 1-1

Description

[Technical Field]

[0001] Related Applications This application claims priority to U.S. Provisional Patent Application No. 62 / 554,271, filed September 5, 2017, and U.S. Provisional Patent Application No. 62 / 685,687, filed June 15, 2018, the entire contents of both of which are incorporated herein by reference.

[0002] The present disclosure relates to methods and compositions for treating inflammatory skin diseases using recombinant microorganisms. [Background technology]

[0003] Atopic dermatitis (AD), or eczema, is a chronic, pruritic, inflammatory skin disease that affects 5–20% of children worldwide (Williams, H. et al., J Allergy Clin. Immunol. 1999;103(1 Pt 1):125–138) and is also frequently found in many adults. The prevalence of atopic dermatitis is increasing, with some forms of atopic dermatitis affecting 11% of the US population (Shaw, TE et al., J Invest Dermatol. 2011;131(1):67–73), or approximately 35 million people, and costing the US alone $5 billion in direct costs. The primary feature of the disease is dry, scaly, itchy skin. Despite the increasing prevalence of atopic dermatitis worldwide and its significant disease burden, few targeted, effective treatment options are available. In particular, many commonly used treatment methods include a wide range of nonspecific approaches, including, but not limited to, skin hydration, bathing, UV treatment, dietary intervention, antibacterial agents, antihistamines, systemic immunomodulators, and topical corticosteroids. However, despite these numerous options, few result in long-lasting symptom resolution, and recurrence of atopic dermatitis is common in many individuals. Furthermore, the 2013 National Health and Wellness Survey revealed significant associated burdens for atopic dermatitis patients, who reported higher levels of healthcare resources (healthcare provider / ER visits), lower health-related quality of life, and nearly twice as much lost work productivity compared with non-atopic dermatitis patients. Furthermore, patients with atopic dermatitis had significantly higher prevalence of allergies (46% vs. 20%), asthma (22% vs. 8%), anxiety (43% vs. 21%), and depression (37% vs. 21%) (Whiteley, J. et al., Current Medical Research and Opinion. 2016:1-32). Thus, there is a significant unmet need to consider the significant burden that atopic dermatitis has on our healthcare system.

[0004] Recent studies have elucidated the pathophysiology of atopic dermatitis, demonstrating that a defective skin barrier is often the primary cause of atopic dermatitis initiation, resulting in both increased transepidermal water loss (TEWL or TWL) and increased antigen and pathogen exposure. At the same time, atopic dermatitis is often characterized by a lack of diversity in the skin microbiota, dominated by Staphylococcus aureus during atopic dermatitis flares and in untreated skin, and by dysbiosis (or microbial imbalance, the severity of which correlates with disease severity; Kong, HH, et al., Genome research. 2012;22(5):850-859; and Figure 1), a hallmark of atopic dermatitis. Finally, there is an activated inflammatory response, driven by IL-4 / IL-13, with a dominant type 2 helper T cell profile (TH2), present in both lesional and non-lesional skin, indicating a systemic switch to a TH2-weighted profile (Sidbury, R. et al., Current allergy and asthma reports. 2017;17(7):42). Thus, an ideal atopic dermatitis treatment would simultaneously address all of these underlying causes: defective skin barrier, dysbiosis, and activated cutaneous immune responses. The present invention addresses these causes.

[0005] Engineered probiotics are a novel approach based on harnessing the skin microbiota for therapeutic purposes. Engineered probiotics, in particular, have significant advantages over other methods of drug delivery because they establish a home on a patient's skin and deliver therapeutic proteins continuously and stably in situ. Furthermore, certain strains of Staphylococcus epidermidis (SE) have demonstrated significant beneficial immunomodulatory and antipathogenic effects in the skin, which correlate with the disease phenotype and severity of atopic dermatitis. Furthermore, delivery of filaggrin, a structural protein derived from profilaggrin, further enhances therapeutic approaches due to filaggrin's role in the skin barrier and its ability to reduce transepidermal water loss and improve skin hydration. The present invention has the surprising advantage of providing methods and compositions for treating skin diseases, such as atopic dermatitis, using genetically engineered recombinant strains of Staphylococcus epidermidis as a dermal drug delivery system that secretes human filaggrin (e.g., AZT-01) to address the pathophysiology of atopic dermatitis. Once applied to the skin, stable colonization of the skin and subsequent secretion of filaggrin in situ can resolve the disease. Benefits of the present invention include its safety as a non-steroidal treatment option, its efficacy due to the present invention's combination of the benefits from the secretion of filaggrin with the benefits of topical application of S. epidermidis, and its ability to be therapeutically effective even with infrequent application (once a day or less). [Prior art documents] [Non-patent literature]

[0006] [Non-Patent Document 1] Williams, H. et al. J Allergy Clin. Immunol. 1999;103(1 Pt 1):125-138 [Non-patent document 2] Shaw, TE et al., J Invest Dermatol. 2011;131(1):67-73 [Non-patent document 3] Whiteley, J. et al., Current Medical Research and Opinion. 2016:1-32 [Non-patent document 4] Kong, HH et al., Genome research. 2012;22(5):850-859 [Non-Patent Document 5] Sidbury, R. et al. Current allergy and asthma reports. 2017;17(7):42 Summary of the Invention

[0007] The present invention therefore addresses a long-felt need for effective treatments for inflammatory skin diseases such as atopic dermatitis. The present invention also represents one of the first reported demonstrations of commensal skin bacteria capable of secreting therapeutic proteins that treat skin diseases.

[0008] The present invention refers to methods and compositions for treating inflammatory skin diseases that comprise an engineered microorganism capable of expressing a therapeutically relevant recombinant fusion polypeptide (i.e., protein, peptide, or amino acid) as an active ingredient.

[0009] In a first aspect, the present invention features a recombinant microorganism capable of secreting a polypeptide, the recombinant microorganism comprising an expression vector including a first coding sequence comprising a gene capable of expressing the polypeptide and a second coding sequence comprising a gene capable of expressing a cell-penetrating peptide. In a related embodiment, the recombinant microorganism further comprises a third coding sequence comprising a gene capable of expressing an export signal. In yet another embodiment, expression of the first coding sequence, the second coding sequence, and the third coding sequence is under the control of a promoter. In another embodiment, the first coding sequence, the second coding sequence, and the third coding sequence are arranged in frame. In yet another related embodiment, the first coding sequence, the second coding sequence, and the third coding sequence are operably linked to a promoter. In one embodiment, the recombinant microorganism is a bacterium or a combination of bacteria. In another embodiment, the polypeptide is filaggrin or a variant (mutant) thereof. In other embodiments, the microorganism is selected from the group consisting of Bifidobacterium, Brevibacterium, Propionibacterium, Lactococcus, Streptococcus, Staphylococcus, Lactobacillus, Enterococcus, Pediococcus, Leuconostoc, or Oenococcus, or a combination thereof. In yet other embodiments, the recombinant microorganism is Staphylococcus epidermidis. In another embodiment, the microorganism secretes a filaggrin fusion protein. In one embodiment, the polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8 and SEQ ID NO:9.In one embodiment, the polypeptide has at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 1. In one embodiment, the polypeptide has at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO:2. In one embodiment, the polypeptide has at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 3. In one embodiment, the polypeptide has at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO:4. In one embodiment, the polypeptide has at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 5. In one embodiment, the polypeptide has at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO:6. In one embodiment, the polypeptide has at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO:7.In one embodiment, the polypeptide has at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 8. In one embodiment, the polypeptide has at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO:9.

[0010] In a further aspect, the invention features a method for producing a live biotherapeutic composition, the method comprising: (a) transfecting a cell with (i) a first coding sequence comprising a nucleic acid sequence capable of expressing a therapeutic polypeptide and (ii) a second coding sequence comprising a nucleic acid sequence capable of expressing a cell-penetrating peptide; (b) causing the transfected cell to produce a therapeutic polypeptide fusion protein; and (c) obtaining the live biotherapeutic composition. In a related embodiment, the method further comprises (iii) transfecting the cell with a third coding sequence comprising a nucleic acid sequence capable of expressing an export signal. In another embodiment, the first coding sequence, the second coding sequence, and the third coding sequence are disposed in a single plasmid. In yet another embodiment, the first coding sequence, the second coding sequence, and the third coding sequence are operably linked to a promoter. In other embodiments, the cell is selected from the group consisting of a microorganism selected from the group consisting of Bifidobacterium, Brevibacterium, Propionibacterium, Lactococcus, Streptococcus, Staphylococcus, Lactobacillus, Enterococcus, Pediococcus, Leuconostoc, or Oenococcus, or a combination thereof. In yet other embodiments, the cell is Staphylococcus epidermidis. In other embodiments, the therapeutic polypeptide fusion protein is a filaggrin fusion protein or variant thereof.

[0011] In another aspect, the invention features a nucleic acid comprising a nucleic acid sequence that encodes a polypeptide described in any one of the aspects or embodiments herein.

[0012] In a further aspect, the invention features a composition obtainable by any one of the methods disclosed or described herein. In related embodiments, the composition includes a pharmaceutically acceptable carrier selected from the group consisting of an aqueous solution, an emulsion, a cream, a lotion, a gel, or an ointment.

[0013] In a further aspect, the present invention features a live biotherapeutic composition comprising a recombinant microorganism comprising: (i) a first coding sequence comprising a nucleic acid sequence capable of expressing a therapeutic polypeptide; (ii) a second coding sequence comprising a nucleic acid sequence capable of expressing a cell-penetrating peptide; (iii) a third coding sequence comprising a nucleic acid sequence capable of expressing an export signal; and (iv) a promoter operably linked to the first coding sequence, the second coding sequence, and the third coding sequence, wherein the first coding sequence, the second coding sequence, and the first coding sequence are capable of expressing a filaggrin fusion product or a variant thereof. In a related embodiment, the recombinant microorganism is Staphylococcus epidermidis. In a further embodiment, the export signal transports the filaggrin fusion product or a variant thereof out of the recombinant microorganism. In yet another embodiment, the cell-penetrating peptide facilitates (promotes) entry of the filaggrin fusion product or a variant thereof into human keratinocytes. In another embodiment, the composition comprises a pharmaceutically acceptable carrier selected from the group consisting of an aqueous solution, emulsion, cream, lotion, gel, or ointment. The invention, in a further aspect, features a kit including any one of the compositions disclosed or described herein and instructions for use.

[0014] In a further aspect, the invention features a method of treating a skin disease, comprising administering to a subject in need thereof any one of the compositions disclosed or described herein. In another embodiment, the skin disease is atopic dermatitis. [Brief explanation of the drawings]

[0015] [Figure 1-1]Figures 1A-F depict the relationship between staphylococcal species and atopic dermatitis. Figure 1A shows the longitudinal trend of the mean proportion of S. aureus in atopic dermatitis at the antecubital and popliteal crease (AcPC, n = 12) stratified by no treatment (trt) and intermittent trrt flares. Figure 1B shows the proportion of S. aureus in AcPC and the Shannon diversity index. Partial correlation (adjusted for disease status, AcPC). Figure 1C shows the longitudinal trend of the mean proportion of S. epidermidis in AcPC. [Figure 1-2] Figure 1D shows the correlation between the proportion of S. aureus and the objective SCORAD score for each site (AcPc, volar forearm [Vf], nares [N]). Partial correlation (adjusted for disease status). [Figure 1-3] Figure 1E shows the longitudinal trends in Shannon diversity in atopic dermatitis categorized by no treatment and intermittent treatment flares (n = 12, Ac). Figure 1F shows the microbiome progression hypothesis for atopic dermatitis. (*) Proposed relationship between skin microbial diversity, proportion of Staphylococcus aureus, and shifts in disease severity. [Figure 2-1] Figure 2A depicts a schematic diagram of the S. epidermidis-based protein delivery system as described herein. The construct design includes a promoter, a ribosome binding site (RBS), a translocation signal, a filaggrin expression sequence, and a cell-penetrating peptide sequence. Figure 2B shows the characterization of a specific promoter for tunable control of protein expression (using GFP as a reporter). Figure 2C shows the characterization of a translocation signal for protein export from the SE to the exterior (using GFP as a reporter). [Figure 2-2] FIG. 2D shows Western blot analysis of human filaggrin produced by S. epidermidis, mouse filaggrin produced by S. epidermidis, and whole mouse skin (using anti-mouse filaggrin antibody). [Figure 3]Figures 3A-3D depict the characterization of GFP-producing S. epidermidis in reconstituted human epidermis (RHE). Figure 3A shows the fluorescence and optical wavelength overlay of the RHE at approximately 25 μm, 2 hours after application of S. epidermidis-GFP. Figures 3B-3D show the fluorescence and optical wavelength overlay of the RHE 2 hours after dermaroller application followed by topical application of S. epidermidis-GFP. Depths were 0 μm (Figure 3B), 50 μm (Figure 3C), and 70 μm (Figure 3D). [Figure 4] Figures 4A-4E depict the characterization of SE-GFP colonization in mice. Figures 4A-4C show in vivo two-photon excitation micrographs of mouse skin 3 days after treatment with GFP-expressing S. epidermidis. Figure 4A shows unshaved mouse ear skin at a depth of 25 μm; Figure 4B shows 50 μm, and Figure 4C shows shaved dorsal skin at a depth of 80 μm. Figure 4D shows an optical micrograph of the dorsal skin of a mouse after application of SE-GFP. Figure 4E shows an optical micrograph of the dorsal skin of a mouse after application of SE-GFP. [Figure 5-1] Figures 5A-5K depict the characterization of proteins with and without RMR signal using 50 µg of GFP as a reporter in RHE. Figures 5A-5D show two-photon images of locally applied GFP with (Figures 5C, 5D) or without (Figures 5A, 5B) RMR signal at 30 min (Figures 5A, 5C, 5E, 5G) or 60 min (Figures 5B, 5D, 5F, 5H). Images are compiled Z-stacks projected onto a 2D plane. [Figure 5-2] Figures 5E-5H show 3D surface analysis to examine the depth of protein penetration into the RHE. Figures 5I-5N show confocal images of GFP (Figures 5K, 5N), GFP+RMR (Figures 5J, 5M), or vehicle (Figures 5I, 5L) using optical (Figures 5L-5N) or fluorescent (Figures 5I-5K) wavelengths. [Figure 5-3] (Figures 5I-5N) Confocal images of GFP (Figures 5K, 5N), GFP+RMR (Figures 5J, 5M), or vehicle (Figures 5I, 5L) using optical (Figures 5L-5N) or fluorescent (Figures 5I-5K) wavelengths are shown. [Figure 6-1] FIG. 6 depicts the experimental outline of 16S rRNA sequencing. [Figure 6-2] This is a continuation of Figure 6-1. [Figure 7] FIG. 7 is a graph showing the hydrophobicity score as a function of amino acid position for the entire human filaggrin (hFLG) sequence (Uniprot P20930). [Figure 8] FIG. 8 is a graph showing hydrophobicity score as a function of amino acid position for the entire mouse filaggrin (mFLG) sequence (NCBI Reference Sequence: XP017175331.1). [Figure 9] FIG. 9 is a graph showing hydrophobicity score as a function of amino acid position for the hFLG region starting at amino acid 1400 through 1800, including unit segments of domains 9 and 10. [Figure 10] FIG. 10 is a graph showing the hydrophobicity score as a function of amino acid position for hFLG[9-10] (1429-1774) from amino acid position 1400 to amino acid position 1800 in hFLG. [Figure 11] FIG. 11 is a graph showing the hydrophobicity score as a function of amino acid position for the start and end positions of hFLG[9-10](1429-1777). [Figure 12] FIG. 12 is a graph showing the hydrophobicity score as a function of amino acid position from the least hydrophobic point to the next least hydrophobic point (indicated by the arrows). [Figure 13-1] Figure 13 shows the alignment of human filaggrin dimers hFLG[3-4], hFLG[5-6], hFLG[7-8], hFLG[9-10], hFLG[11-12], hFLG[13-14], hFLG[15-16], hFLG[17-18], hFLG[19-20], and hFLG[21-22]. [Figure 13-2] This is a continuation of Figure 13-1. [Figure 13-3] This is a continuation of Figure 13-1. [Figure 14]FIG. 14 is a graph showing background FLG binding at 1 μg / well over 2 h at 37° C. (non-specific binding—NSB). [Figure 15] FIG. 15 is a graph showing binding of hFLG segments to human callus keratin (NSB removed). [Figure 16] FIG. 16 is a graph showing titration of IgY anti-hFLG. DETAILED DESCRIPTION OF THE INVENTION

[0016] I. Overview and Definitions Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this invention belongs.The following references provide the general definitions of many terms used in this invention: Singleton et al., Dictionary of Microbiology and Molecular Biology (2nd edition, 1994); The Cambridge Dictionary of Science and Technology (Walker (ed.), 1988); The Glossary of Genetics, 5th edition, R. Rieger et al. (ed.), Springer Verlag (1991); and Hale & Marham, The HarperCollins Dictionary of Biology (1991).As used herein, the following terms have the meanings ascribed to them unless otherwise specified.

[0017] The articles "a" and "an" are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. For example, "an element" means one element or more than one element.

[0018] As used herein, the term "including" means, and is used interchangeably with, the phrase "including but not limited to."

[0019] As used herein, the term "or" means, and is used interchangeably with, the term "and / or," unless context clearly indicates otherwise.

[0020] As used herein, the term "such as" means, and is used interchangeably with, the phrase "such as but not limited to."

[0021] As used herein, the following terms have the following meanings unless expressly stated to the contrary. As used herein, the term "abnormal skin condition" or "skin disease" (e.g., inflammatory skin disease) refers to a skin condition or condition that is generally undesirable or harmful compared to the normal or baseline condition of human skin. Examples of abnormal skin conditions include psoriasis, acne, atopic dermatitis, allergic contact dermatitis, epidermolytic hyperkeratosis, seborrheic dermatitis, eczema, dry skin, allergies, rashes, UV-irritated skin, detergent-irritated skin (including irritation caused by enzymes and molecules used in detergents and sodium lauryl sulfate), thinning skin (e.g., skin from the elderly and children), bullous pemphigoid, pemphigus vulgaris, impetigo, vitiligo, alopecia, and hirsutism.

[0022] As used herein, the term "patient" or "subject" refers to a human or animal (in the case of animals, more typically mammals, such as domesticated mammals, or poultry animals and animals such as fish and other marine or freshwater food organisms) that will receive the treatments and compositions of the present invention. Such patients or subjects would be in need of the pharmaceutical compositions of the present invention or methods for treating, preventing, or reducing the risk of abnormal skin conditions or skin diseases (e.g., inflammatory skin diseases).

[0023] As used herein, the term "therapeutically effective amount" refers to the amount of a pharmaceutically active compound, live biotherapeutic composition, combination of compounds or compositions, or the amount of pharmaceutically active compound delivered by an engineered bacterial strain or strains, e.g., a skin treatment agent or agents, when administered alone or in combination to treat, prevent, or reduce the risk of a disease state or condition, such as an abnormal skin condition or skin disease (e.g., inflammatory skin disease). The term also refers to the amount of an active compound or combination of compounds or a pharmaceutical composition containing an engineered bacterial strain or strains that delivers a pharmaceutically active compound. For example, an effective amount refers to the amount of compound or the amount of compound delivered by an engineered bacterial strain (or recombinant bacterial strain) or strains present in a formulation given to a recipient patient or subject sufficient to elicit biological activity, e.g., activity to treat or prevent an abnormal skin condition or skin disease (e.g., inflammatory skin disease).

[0024] As used herein, the phrase "pharmaceutically acceptable" refers to active compounds, materials, engineered bacterial strain or strains, compositions, carriers, and / or dosage forms that are suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, within the scope of sound medical judgment, commensurate with a reasonable benefit / risk ratio. As used herein, the term "treating" refers to providing a therapeutic intervention to cure or ameliorate an abnormal skin condition.

[0025] As used herein, the term "preventing" refers to completely or nearly completely stopping the occurrence of an abnormal skin condition, for example, when a patient or subject is predisposed to or at risk of developing an abnormal skin condition. Preventing may also include inhibiting, i.e., stopping, the occurrence of an abnormal skin condition.

[0026] As used herein, the term "reducing risk" refers to reducing the likelihood or probability of an abnormal skin condition occurring, for example, when a patient or subject is predisposed to or at risk of suffering from an abnormal skin condition.

[0027] As used herein, the term "engineered bacterial strain" or "recombinant bacterial strain" refers to a bacterial strain that has been "genetically modified" or "engineered" by introducing DNA prepared outside the organism into the bacterial strain. For example, introducing a plasmid containing new genes or other nucleic acid sequences into a bacterium can cause the bacterium to express those genes or other nucleic acid sequences. Alternatively, after introducing the plasmid containing the new genes or other nucleic acid sequences into the bacterium, it can be integrated into the bacterial genome, where the bacterium will express those genes or other nucleic acid sequences.

[0028] As used herein, the term "carrier," "carrier system," or "vehicle" refers to a compatible substance suitable for delivering, containing, or "carrying" an active pharmaceutical ingredient or other material for administration in a composition topically applied to a patient or subject. Carriers useful herein should be pharmaceutically acceptable. Carriers and vehicles useful herein include any such material known in the art that is non-toxic and does not adversely interact with other components of the formulation in which it is contained. The term "aqueous" refers to a formulation that contains water or becomes water-containing after application to the skin or mucosal tissue. Further examples of "carriers" include water, lower alcohols, higher alcohols, polyhydric alcohols, monosaccharides, disaccharides, polysaccharides, hydrocarbon oils, fats and oils, waxes, fatty acids, silicone oils, nonionic surfactants, ionic surfactants, silicone surfactants, and water-based and emulsion-based mixtures of such carriers.

[0029] As used herein, the term "polypeptide" or "protein" refers to a biological molecule, or macromolecule, made up of amino acid residues linked together in a chain. The definition of polypeptide as used herein is intended to encompass proteins (generally higher molecular weight) made up of one or more long chains of amino acid residues, as well as small molecules (generally lower molecular weight) of a few amino acids. In other embodiments, single amino acids, although not technically polypeptides, are also considered within the scope of the present invention.

[0030] As used herein, the term "live biotherapeutic product" (or LBP) refers to a product candidate that contains bacteria, yeast, and / or other microorganisms.

[0031] For purposes of the present invention, the term "isolated" refers to biological material (cells, nucleic acids, or proteins) that has been removed from its original environment (the environment in which it naturally occurs). For example, a polynucleotide that occurs in the natural state in a plant or animal is not isolated, but the same polynucleotide separated from the adjacent nucleic acids with which it naturally occurs is considered to be "isolated."

[0032] An "isolated nucleic acid molecule" (e.g., an isolated promoter) is one that is separated from other nucleic acid molecules that are present in the natural source of the nucleic acid. For example, with respect to genomic DNA, the term "isolated" includes a nucleic acid molecule that is separated from the chromosome with which the genomic DNA is naturally associated. Preferably, an "isolated" nucleic acid molecule is free of sequences that naturally flank the nucleic acid molecule in the genomic DNA of the organism from which the nucleic acid molecule is derived.

[0033] The present invention provides skin-colonizing bacteria that have been genetically altered to express a recombinant therapeutic polypeptide for the treatment or prevention of skin diseases (Figure 2). The use of genetically engineered protein-producing bacteria has several advantages over prior art methods for treating skin diseases. The therapeutic protein can treat the underlying cause of the defect that results in the skin condition. Furthermore, the bacteria can replicate autonomously while retaining the inserted gene to continuously produce the therapeutic protein.

[0034] The present invention provides skin-colonizing bacteria, such as Staphylococcus epidermidis, that have been genetically altered to express human filaggrin. The use of genetically engineered filaggrin-producing bacteria has several advantages over the use of filaggrin supplementation. First, the bacteria can self-replicate while retaining the inserted filaggrin gene. Second, Staphylococcus epidermidis has been shown to inhibit the growth of Staphylococcus aureus, a bacterial species normally present on the skin that dominates the skin microflora during AD flare-ups.

[0035] II. Methods and Compositions of the Invention The present invention provides skin-colonizing microorganisms, e.g., bacteria, that have been genetically altered to express a recombinant therapeutic polypeptide for the treatment or prevention of skin diseases (Figure 2). The use of genetically engineered protein-producing microorganisms, e.g., bacteria, has several advantages over prior art methods for treating skin diseases. The therapeutic protein can treat the underlying cause of the skin condition. Furthermore, the microorganisms, e.g., bacteria, can self-replicate while retaining the inserted nucleic acid (e.g., gene) to continuously produce the therapeutic protein.

[0036] The present invention provides a skin-colonizing microorganism, e.g., a bacterium, such as Staphylococcus epidermidis, that has been genetically altered to express a therapeutic protein, e.g., human filaggrin. The use of genetically engineered filaggrin-producing microorganisms, e.g., bacteria, has several advantages over the use of filaggrin supplementation. First, the microorganism, e.g., bacteria, can self-replicate while retaining the inserted filaggrin nucleic acid sequence (e.g., gene). Second, Staphylococcus epidermidis has been shown to inhibit the growth of Staphylococcus aureus, a bacterial species normally present on the skin that dominates the skin microflora during atopic dermatitis flare-ups.

[0037] Bacterial strains The present invention provides genetically altered microorganisms, such as bacteria, capable of expressing recombinant therapeutic proteins. A wide range of microorganisms are suitable for use in the present invention. Examples include, but are not limited to, non-pathogenic and commensal bacteria. Bacteria suitable for use in the present invention include, but are not limited to, Bifidobacterium, Brevibacterium, Propionibacterium, Lactococcus, Streptococcus, Staphylococcus (e.g., S. epidermidis), Lactobacillus (e.g., L. acidophilus), Pediococcus, Leuconostoc, or Oenococcus. In certain embodiments of the present invention, the bacterium is Staphylococcus epidermidis. In preferred embodiments of the present invention, the strain of Staphylococcus epidermidis used is incapable of producing biofilms. One such example of a strain of Staphylococcus epidermidis incapable of producing biofilms is the Staphylococcus epidermidis strain ATCC 12228. However, in still other embodiments of the present invention, other related or similar species found on the skin may be used.

[0038] Therapeutic Proteins The present invention provides genetically altered microorganisms, such as bacteria, capable of expressing recombinant therapeutic proteins.

[0039] In some embodiments, the present disclosure relates to a therapeutic protein comprising a filaggrin polypeptide amino acid sequence. In some embodiments, the present disclosure relates to a therapeutic protein comprising a filaggrin polypeptide amino acid sequence and a cell-penetrating polypeptide amino acid sequence. In some embodiments, the present disclosure relates to a therapeutic protein comprising a filaggrin polypeptide amino acid sequence, a cell-penetrating polypeptide amino acid sequence, and a secretory or export signal polypeptide sequence. As used herein, "polypeptide" is generally defined herein to refer to a peptide sequence of about 2 to about 10,000 or more amino acid residues. The term "amino acid" encompasses not only the 20 common amino acids found in naturally synthesized proteins, but also any modified, unconventional, or synthetic amino acids. Those of skill in the art will be familiar with modified, unconventional, or synthetic amino acids.

[0040] The polypeptides of the present invention may have amino acid deletions and / or substitutions compared to the native sequence; thus, sequences with deletions, sequences with substitutions, and sequences with deletions and substitutions are contemplated (envisioned) for inclusion in the polypeptides of the present invention. In some embodiments, these polypeptides may further comprise inserted or additional amino acids, e.g., linkers.

[0041] Substitution or replacement variants typically contain the exchange of one amino acid for another at one or more sites within the protein and can be designed to modulate one or more properties of the polypeptide, particularly, to increase its potency or specificity. Substitutions of this type preferably are conservative, i.e., one amino acid is replaced with one of similar shape and charge. Conservative substitutions are well known in the art, and include, for example, alanine to serine; arginine to lysine; asparagine to glutamine or histidine; aspartic acid to glutamic acid; cysteine ​​to serine; glutamine to asparagine; glutamic acid to aspartic acid; glycine to proline; histidine to asparagine or glutamine; isoleucine to leucine or valine; leucine to valine or isoleucine; lysine to arginine; methionine to leucine or isoleucine; phenylalanine to tyrosine, leucine, or methionine; serine to threonine; threonine to serine; tryptophan to tyrosine; tyrosine to tryptophan or phenylalanine; and valine to isoleucine or leucine. In addition to deletion or substitution, a polypeptide may have one or more residue insertions. This may include the addition of one or more amino acid residues.

[0042] Amino acid substitutions are generally based on the relative similarity of the amino acid side-chain substituents, for example, their hydrophobicity, hydrophilicity, charge, size, etc. Exemplary substitutions taking various such characteristics into consideration are well known to those of skill in the art and include: arginine for lysine; glutamic acid for aspartic acid; serine for threonine; glutamic acid for aspartic acid; and valine, leucine, and isoleucine.

[0043] Filaggrin In some embodiments, the present disclosure relates to therapeutic proteins comprising a filaggrin polypeptide amino acid sequence. In preferred embodiments of the present invention, the therapeutic protein comprises human filaggrin. Human filaggrin is expressed by a human gene encoding filaggrin (FLG). Filaggrin is a protein produced by differentiating keratinocytes and functions to aggregate keratin filaments, along with other components, into the cytoskeleton that comprises the keratinocyte envelope. FLG is a large gene located on chromosome lq21 that produces profilaggrin, an insoluble polyprotein that is proteolyzed to release functional filaggrin monomers (Armengot-Carbo et al., 2014). The therapeutic proteins of the present invention (and thus the genes from which the proteins are expressed) may be derived from any mammal. Non-limiting examples include, but are not limited to, mouse, rat, rabbit, goat, sheep, horse, cow, dog, primate, or human gene sequences.

[0044] The filaggrin amino acid sequence intended for inclusion in the polypeptide, composition, and method of the present invention can be obtained from any source.For example, filaggrin amino acid can be obtained from natural sources or chemically synthesized.The filaggrin amino acid sequence can be derived from any species.For example, it can be a mammalian filaggrin amino acid sequence.Non-limiting examples include mouse, rat, rabbit, goat, sheep, horse, cow, dog, cat, primate, or human amino acid sequence.In a preferred embodiment, the filaggrin amino acid sequence is a human amino acid sequence.Non-limiting examples of filaggrin proteins are listed in Table 1 below.

[0045] [Table 1]

[0046] In some embodiments, the filaggrin amino acid sequence comprises any of the amino acid sequences set forth in Table 1. In certain embodiments, the filaggrin amino acid sequence comprises GenBank Accession No. NP_002007.1 (SEQ ID NO: 1).

[0047] In some embodiments, the filaggrin amino acid sequence comprises 20, 30, 40, 50, 60, 70, 80, 90, 100, 120, 130, 150, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340 or more consecutive amino acids of any of the amino acid sequences set forth in Table 1, so long as the filaggrin amino acid sequence, when conjugated to a cell-penetrating peptide and / or translocation or secretion signal, retains at least some of the function of a native filaggrin amino acid sequence conjugated to the same cell-penetrating peptide and / or translocation or secretion signal.

[0048] In some embodiments, the filaggrin amino acid sequence has at least about 80%, 81%, 82%, 83, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to a native filaggrin amino acid sequence, or any range of percent sequence identity derivable therein. In one embodiment, the filaggrin amino acid sequence is an amino acid sequence selected from Table 1. In some embodiments, the filaggrin amino acid sequence has at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO:1.

[0049] In some embodiments, the filaggrin amino acid sequence has at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO:2.

[0050] In some embodiments, the filaggrin amino acid sequence has at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO:3.

[0051] In some embodiments, the filaggrin amino acid sequence has at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO:4.

[0052] In some embodiments, the filaggrin amino acid sequence has at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO:5.

[0053] In some embodiments, the filaggrin amino acid sequence has at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO:6.

[0054] In some embodiments, the filaggrin amino acid sequence has at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO:7.

[0055] In some embodiments, the filaggrin amino acid sequence has at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO:8.

[0056] In some embodiments, the human filaggrin consensus sequence is the consensus sequence shown as SEQ ID NO: 9, which refers to a sequence formed from the amino acids most frequently present in hFLG[3-4], hFLG[5-6], hFLG[7-8], hFLG[9-10], hFLG[11-12], hFLG[13-14], hFLG[15-16], hFLG[17-18], hFLG[19-20], and hFLG[21-22]. SEQ ID NO:9 JPEG2024009822000003.jpg63163

[0057] In some embodiments, the filaggrin amino acid sequence has at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO:9.

[0058] "Sequence identity" is defined as the percentage of amino acid residues in a candidate sequence that are identical to the amino acid residues at the corresponding positions in a native polypeptide sequence after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity without considering any conservative substitutions as part of the sequence identity. The sequence identity percentage value can be generated by the NCBI BLAST 2.0 software as defined by Altschul et al. (1997). Parameters are set to default values, except for the penalty for mismatches, which is set to -1.

[0059] In a preferred embodiment of the invention, the therapeutic protein comprises a recombinant fusion protein comprising filaggrin operably linked to a cell-penetrating protein (CPP). In another embodiment of the invention, the therapeutic protein comprises a recombinant fusion protein comprising filaggrin operably linked to a translocation or secretion signal capable of transporting the recombinant filaggrin to the exterior of a microorganism (e.g., a bacterium). In another embodiment, the therapeutic protein comprises a recombinant fusion protein comprising filaggrin operably linked to a cell-penetrating protein (CPP) and a translocation or secretion signal.

[0060] Furthermore, the polypeptides described herein may contain a sequence of any number of additional amino acid residues at either the N-terminus or C-terminus of the amino acid sequence comprising the filaggrin amino acid sequence and the cell-penetrating protein (CPP) and / or the translocation or secretion signal. For example, an amino acid sequence of about 3 to about 10,000 or more amino acid residues may be present at either the N-terminus, C-terminus, or both the N-terminus and C-terminus of the amino acid sequence comprising the filaggrin amino acid sequence and the cell-penetrating peptide and / or the translocation or secretion signal.

[0061] secretion signal A secretion signal or export signal is a peptide sequence on a protein that facilitates the transport of the protein through the secretory pathway, ultimately resulting in the secretion of the protein from the cell. In the present invention, any secretion signal that facilitates the transport of a protein, such as a filaggrin-containing protein, from a microorganism (e.g., a bacterial cell) to the outside is contemplated as a secretion signal.

[0062] Cell-penetrating peptides A cell-penetrating peptide is a peptide sequence that facilitates or mediates the delivery of a biomolecule (e.g., a protein) in vivo without using a receptor and without causing significant membrane damage. Cell-penetrating peptides that facilitate entry into skin keratinocytes are contemplated as cell-penetrating peptides of the present invention.

[0063] According to some embodiments, the present disclosure provides a recombinant microorganism capable of secreting a polypeptide comprising a filaggrin polypeptide amino acid sequence, the recombinant microorganism comprising an expression vector comprising a first coding sequence comprising a gene capable of expressing the polypeptide and a second coding sequence comprising a gene capable of expressing a cell-penetrating peptide.

[0064] According to some embodiments, the present disclosure provides a recombinant microorganism capable of secreting a polypeptide comprising an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:1, the recombinant microorganism comprising an expression vector comprising a first coding sequence comprising a gene capable of expressing the polypeptide and a second coding sequence comprising a gene capable of expressing a cell-penetrating peptide.

[0065] According to some embodiments, the present disclosure provides a recombinant microorganism capable of secreting a polypeptide comprising an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:2, the recombinant microorganism comprising an expression vector comprising a first coding sequence comprising a gene capable of expressing the polypeptide and a second coding sequence comprising a gene capable of expressing a cell-penetrating peptide.

[0066] According to some embodiments, the present disclosure provides a recombinant microorganism capable of secreting a polypeptide comprising an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:3, the recombinant microorganism comprising an expression vector comprising a first coding sequence comprising a gene capable of expressing the polypeptide and a second coding sequence comprising a gene capable of expressing a cell-penetrating peptide.

[0067] According to some embodiments, the present disclosure provides a recombinant microorganism capable of secreting a polypeptide comprising an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:4, the recombinant microorganism comprising an expression vector comprising a first coding sequence comprising a gene capable of expressing the polypeptide and a second coding sequence comprising a gene capable of expressing a cell-penetrating peptide.

[0068] According to some embodiments, the present disclosure provides a recombinant microorganism capable of secreting a polypeptide comprising an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:5, the recombinant microorganism comprising an expression vector comprising a first coding sequence comprising a gene capable of expressing the polypeptide and a second coding sequence comprising a gene capable of expressing a cell-penetrating peptide.

[0069] According to some embodiments, the present disclosure provides a recombinant microorganism capable of secreting a polypeptide comprising an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:6, the recombinant microorganism comprising an expression vector comprising a first coding sequence comprising a gene capable of expressing the polypeptide and a second coding sequence comprising a gene capable of expressing a cell-penetrating peptide.

[0070] According to some embodiments, the present disclosure provides a recombinant microorganism capable of secreting a polypeptide comprising an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:7, the recombinant microorganism comprising an expression vector comprising a first coding sequence comprising a gene capable of expressing the polypeptide and a second coding sequence comprising a gene capable of expressing a cell-penetrating peptide.

[0071] According to some embodiments, the present disclosure provides a recombinant microorganism capable of secreting a polypeptide comprising an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:8, the recombinant microorganism comprising an expression vector comprising a first coding sequence comprising a gene capable of expressing the polypeptide and a second coding sequence comprising a gene capable of expressing a cell-penetrating peptide.

[0072] According to some embodiments, the present disclosure provides a recombinant microorganism capable of secreting a polypeptide comprising an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:9, the recombinant microorganism comprising an expression vector comprising a first coding sequence comprising a gene capable of expressing the polypeptide and a second coding sequence comprising a gene capable of expressing a cell-penetrating peptide.

[0073] According to some embodiments, the present disclosure provides a recombinant microorganism capable of secreting a polypeptide, the recombinant microorganism comprising an expression vector comprising an amino acid sequence that is at least 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:2.

[0074] According to some embodiments, the present disclosure provides a recombinant microorganism capable of secreting a polypeptide, the recombinant microorganism comprising an expression vector comprising an amino acid sequence that is at least 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:3.

[0075] According to some embodiments, the present disclosure provides a recombinant microorganism capable of secreting a polypeptide, the recombinant microorganism comprising an expression vector comprising an amino acid sequence that is at least 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:4.

[0076] According to some embodiments, the present disclosure provides a recombinant microorganism capable of secreting a polypeptide, the recombinant microorganism comprising an expression vector comprising an amino acid sequence that is at least 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:5.

[0077] According to some embodiments, the present disclosure provides a recombinant microorganism capable of secreting a polypeptide, the recombinant microorganism comprising an expression vector comprising an amino acid sequence that is at least 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:6.

[0078] According to some embodiments, the present disclosure provides a recombinant microorganism capable of secreting a polypeptide, the recombinant microorganism comprising an expression vector comprising an amino acid sequence that is at least 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:7.

[0079] According to some embodiments, the present disclosure provides a recombinant microorganism capable of secreting a polypeptide, the recombinant microorganism comprising an expression vector comprising an amino acid sequence that is at least 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:8.

[0080] nucleic acid The present invention includes nucleic acids comprising nucleic acid sequences encoding the recombinant polypeptides of the present invention. Some embodiments of the present invention include nucleic acids comprising nucleic acid sequences encoding the above polypeptides. Further embodiments include nucleic acids encoding filaggrin amino acid sequences. The filaggrin amino acid sequence is any of the filaggrin amino acid sequences described herein. In some embodiments, the nucleic acid is contained in an expression vector. The term "nucleic acid" is well known in the art. As used herein, "nucleic acid" generally refers to a molecule (i.e., a strand) of DNA, RNA, or a derivative or analog thereof, comprising nucleobases. Nucleobases include, for example, naturally occurring purine or pyrimidine bases found in DNA (e.g., adenine "A," guanine "G," thymine "T," or cytosine "C") or RNA (e.g., A, G, uracil "U," or C). The term "nucleic acid" encompasses the terms "oligonucleotide" and "polynucleotide," respectively, as subgenus of the term "nucleic acid." The term "oligonucleotide" refers to a molecule of 3 to about 100 nucleobases in length. The term "polynucleotide" refers to at least one molecule of greater than about 100 nucleobases in length.

[0081] These definitions refer to single-stranded or double-stranded nucleic acid molecules. Double-stranded nucleic acids are formed by perfectly complementary bonds, although in some embodiments, double-stranded nucleic acids may be formed by partial or substantial complementary bonds. Thus, nucleic acids may encompass double-stranded molecules that typically contain one or more complementary strands or "complements" of a particular sequence comprising a molecule. As used herein, single-stranded nucleic acids may be designated with the prefix "ss" and double-stranded nucleic acids may be designated with the prefix "ds."

[0082] Gene constructs The present invention uses standard molecular biology techniques, such as those described in (Sambrook et al., 2001). An example of a genetic construct used in the present invention is pAZT, an allelic exchange E. coli-Staphylococcus aureus shuttle vector based on pJB38, which further contains additional design features on the plasmid to improve function (Bose, JL, et al. Applied and Environmental Microbiology. 2013;79(7):2218-2224). Plasmids are constructed using standard molecular biology techniques by inserting the cDNA of a gene encoding a therapeutic protein into a restriction site (Figure 2). The insert further contains a coding sequence driven by a promoter. Such promoters may be constitutive or inducible. Examples of inducible promoters include those activated by compounds such as alcohol, sugars, metals, or tetracycline, or by physical factors such as light or high temperature.

[0083] The mRNA sequence of human FLG has the Genebank accession number NM_002016. Plasmid pAZT was constructed by inserting a portion of the FLG cDNA into the restriction site of pJB38. The insert contains a promoter-driven nucleic acid coding sequence. The construct further contains nucleic acid sequences encoding a secretion signal and a cell-penetrating peptide, resulting in a recombinant filaggrin fusion protein.

[0084] Use of recombinant bacterial strains It is understood that the skin disease to be treated may be any disease or disorder related to the skin. In a preferred embodiment, the disorder is selected from the group consisting of atopic dermatitis, psoriasis, acne, allergic contact dermatitis, epidermolytic hyperkeratosis, seborrheic dermatitis, eczema, dry skin, allergies, rash, UV-irritated skin, detergent-irritated skin (including irritation caused by enzymes and compounds used in detergents and sodium lauryl sulfate), thinning skin (e.g., skin from the elderly and children), bullous pemphigoid, pemphigus vulgaris, impetigo, vitiligo, alopecia, and hypertrichosis. Examples of proteins that can be administered according to the present invention are preferably eukaryotic proteins. These proteins include, but are not limited to, single amino acids, small peptides, and large proteins. More specifically, genes encoding proteins useful in the present invention as recombinant therapeutic proteins include, but are not limited to, the following: members of the interleukin family of genes, including, but not limited to, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, IL-13, IL-14, and IL-15, and genes encoding their receptor antagonists. Genes encoding hematopoietic growth factors, including, but not limited to, erythropoietin, granulocyte colony-stimulating factor, granulocyte-macrophage colony-stimulating factor, macrophage colony-stimulating factor, stem cell factor, leukemia inhibitory factor, and thrombopoietin, are also contemplated in the present invention. Genes encoding neurotrophic factors, including, but not limited to, nerve growth factor, brain-derived neurotrophic factor, and ciliary neurotrophic factor, are also contemplated. Additionally, genes encoding interferons, including, but not limited to, IFN-alpha, IFN-beta, and IFN-gamma, are included.Also contemplated by the present invention are genes encoding chemocacins such as cytokines of the CC and CXC families, genes encoding hormones such as proinsulin and growth hormone (growth hormone), and genes encoding thrombolytic enzymes, including other enzymes such as tissue plasminogen activator, streptokinase, urokinase, or trypsin inhibitor. The present invention further includes genes encoding tissue repair, growth, and regulatory factors, including, but not limited to, oncostatin M, platelet-derived growth factor, fibroblast growth factor, epidermal growth factor, hepatocyte growth factor, bone morphogenetic protein, insulin-like growth factor, calcitonin, and transforming growth factor alpha and beta. Further contemplated genes include genes encoding structural proteins, including filaggrin, actin, collagen, fibrillin, elastin, or scleroproteins.

[0085] formulation A formulation for use according to the invention may comprise a pharmaceutically effective amount of a genetically engineered microorganism, e.g., a bacterium, that produces a therapeutically effective amount of a desired polypeptide, e.g., at least about 0.01%, about 0.05%, about 0.1%, about 0.2%, about 0.3%, about 0.4%, about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, about 1.0%, about 1.5%, about 2.0%, about 3.0%, about 4.0%, about 5.0%, about 6.0%, about 7.0%, about 8.0%, about 9.0%, about 10.0%, about 11.0%, about 12.0%, about 13.0%, about 14.0%, about 15.0%, about 16.0%, about 17.0%, about 18.0%, about 19.0%, about 20.0%, about 21.0%, about 22.0%, about 23.0%, about 24.0%, about 25.0%, about 26.0%, about 27.0%, about 28.0%, about 29.0%, about 30.0%, about 31.0%, about 32.0%, about 33.0%, about 34.0%, about 35.0%, about 36.0%, about 37.0%, about 38.0%, about 39.0%, about 40.0%, about 41.0%, about 42.0%, about 43.0%, about 44.0%, about 45.0%, about 46.0%, about 47.0%, about 48.0%, about 49.0%, about 50.0%, about 51.0%, about 52.0%, about 53.0%, about 54.0%, about 55.0%, about 56.0 It will be further apparent that the composition may comprise 0%, about 10.0%, about 11.0%, about 12.0%, about 13.0%, about 14.0%, about 15.0%, about 16.0%, about 17.0%, about 18.0%, about 19.0%, about 20.0%, about 25.0%, about 30.0%, about 35.0%, about 40.0%, about 45.0%, about 50.0% or more by weight of genetically engineered microorganisms, e.g., bacteria, with the upper limit being about 90.0% by weight of genetically engineered microorganisms, e.g., bacteria.

[0086] In alternative embodiments, formulations for use according to the present invention may contain, for example, at least about 0.01% to about 30% by weight, about 0.01% to about 20% by weight, about 0.01% to about 5% by weight, about 0.1% to about 30% by weight, about 0.1% to about 20% by weight, about 0.1% to about 15% by weight, about 0.1% to about 10% by weight, about 0.1% to about 5% by weight, about 0.2% to about 5% by weight, about 0.3% to about 5% by weight, about 0.4% to about 5% by weight, about 0.5% to about 5% by weight, about 1% to about 5% by weight or more of a genetically engineered microorganism, e.g., a bacterium.

[0087] Topical formulations for use in the present invention may be in any form suitable for application to body surfaces, such as creams, lotions, sprays, solutions, gels, ointments, pastes, salves, paints, bioadhesives, suspensions, emulsions, etc., and / or may be prepared to contain liposomes, micelles, and / or microspheres. Such formulations may be used in combination with an occlusive covering layer so that moisture evaporating from the body surface is retained within the formulation during and after application to the body surface. The formulation may include a live biotherapeutic composition or may include at least one genetically engineered microorganism, e.g., an engineered bacterial strain, that produces a recombinant polypeptide. This engineered live biotherapeutic composition can deliver the polypeptide directly to the skin to treat or prevent abnormal skin conditions and / or skin diseases (e.g., inflammatory skin diseases).

[0088] Topical formulations include those in which any other active ingredient is dissolved or dispersed in a dermatological vehicle known in the art, such as an aqueous or non-aqueous gel, an ointment, a water-in-oil or oil-in-water emulsion, etc. Components of such vehicles may include water, an aqueous buffer solution, a non-aqueous solvent (such as ethanol, isopropanol, benzyl alcohol, 2-(2-ethoxyethoxy)ethanol, propylene glycol, propylene glycol monolaurate, glycofurol, or glycerol), an oil (e.g., a mineral oil such as liquid paraffin, a natural or synthetic triglyceride such as MIGLYOL, or a silicone oil such as dimethicone). Depending on the nature of the formulation and its intended use and site of application, the dermatological vehicle used may contain one or more ingredients (e.g., ingredients in addition to water if the formulation is an aqueous gel) selected from the following: solubilizers or solvents (e.g., β-cyclodextrins such as hydroxypropyl β-cyclodextrin, or alcohols or polyols such as ethanol, propylene glycol, or glycerol); thickeners (e.g., hydroxyethyl cellulose, hydroxypropyl cellulose, carboxymethyl cellulose, or carbomer); gelling agents (e.g., polyoxyethylene-polyoxypropylene copolymers); preservatives (e.g., benzyl alcohol, benzalkonium chloride, chlorhexidine, chlorbutol, benzoates, potassium sorbate, or EDTA or salts thereof); and pH buffers (such as mixtures of dihydrogen phosphate and hydrogen phosphate, or mixtures of citric acid and hydrogen phosphate).

[0089] Pharmaceutically acceptable carriers may be included in the formulations of the present invention, and may be any carrier conventionally used in the art.Examples include water, lower alcohols, higher alcohols, polyhydric alcohols, monosaccharides, disaccharides, polysaccharides, hydrocarbon oils, fats and oils, waxes, fatty acids, silicone oils, nonionic surfactants, ionic surfactants, silicone surfactants, and water-based and emulsion-based mixtures of such carriers.The term "pharmaceutically acceptable" or "pharmaceutically acceptable carrier" used herein refers to a compound or composition that can be included in a pharmaceutical formulation without causing undesired biological effects or undesired interactions with other components of the formulation, and "carrier" or "vehicle" used herein refers to a carrier material that is suitable for incorporation into a topically applied composition.The carriers and vehicles useful herein include any such materials known in the art that are non-toxic and do not interact with other components of the formulation in a harmful manner.The term "aqueous" refers to a formulation that contains water or becomes water-based after application to skin or mucous membrane tissue.

[0090] Cream bases are water-washable and contain an oil phase, an emulsifier, and an aqueous phase. The oil phase, also called the "internal" phase, typically contains petrolatum and a fatty alcohol such as cetyl or stearyl alcohol. The aqueous phase typically, but not necessarily, exceeds the oil phase in volume and typically contains a moisturizer. The emulsifier in a cream formulation is typically a nonionic, anionic, cationic, or amphoteric surfactant.

[0091] Lotions are preparations that are applied to the skin surface without friction, and are typically liquid or semi-liquid preparations in which particles containing active agents are present in a water or alcohol base. Lotions are usually suspensions of solids, preferably containing a liquid oily emulsion of the oil-in-water type. Lotions are the preferred formulations herein for treating large body areas because they allow for easier application of more liquid compositions. It is generally necessary for insoluble substances in lotions to be finely divided. Lotions typically contain suspending agents to provide better dispersion, as well as compounds useful for localizing and retaining active agents in contact with the skin, such as methylcellulose, sodium carboxymethylcellulose, etc. A solution is a homogeneous mixture prepared by dissolving one or more chemicals (solutes) in a liquid so that the molecules of the dissolved substance are dispersed among the molecules of the solvent. A solution may contain other pharmaceutically or cosmetically acceptable chemicals to buffer, stabilize, or retain the solute. Common examples of solvents used in preparing solutions are ethanol, water, propylene glycol, or any other acceptable vehicle. As is well known, gels are semisolid, suspension-type systems. Single-phase gels typically contain organic polymers substantially uniformly distributed throughout a carrier liquid, which is aqueous, but preferably also contains alcohol and, optionally, oil. Preferred organic polymers, i.e., gelling agents, are crosslinked acrylic acid polymers, such as the "carbomer" family of polymers, e.g., carboxypolyalkylenes commercially available under the trademark Carbopol. Also preferred are hydrophilic polymers such as polyethylene oxide, polyoxyethylene-polyoxypropylene copolymers, and polyvinyl alcohol; cellulosic polymers such as hydroxypropyl cellulose, hydroxyethyl cellulose, hydroxypropylmethylcellulose, hydroxypropylmethylcellulose phthalate, and methylcellulose; gums such as tragacanth and xanthan gum; sodium alginate; and gelatin.To prepare a uniform gel, a dispersant such as alcohol or glycerin can be added, or the gelling agent can be dispersed by grinding, mechanical mixing, stirring, or a combination thereof. Ointments, also well known in the art, are typically semi-solid preparations based on petrolatum or other petroleum derivatives. The specific ointment base used will provide several desirable characteristics, such as emollient properties, as will be understood by those skilled in the art. Like other carriers or vehicles, ointment bases should be inert, stable, non-irritating, and non-sensitizing. As explained in Remington: The Science and Practice of Pharmacy, 19th Edition (Easton, PA: Mack Publishing Co., 1995), pages 1399-1404, ointment bases can be classified into four classes: fatty bases; emulsifiable bases; emulsion bases; and water-soluble bases. Fatty ointment bases include, for example, vegetable oils, animal fats, and semi-solid hydrocarbons derived from petroleum.

[0092] Emulsifiable ointment bases, also known as absorbent ointment bases, contain little or no water and include, for example, hydroxystearin sulfate, anhydrous lanolin, and hydrophilic petrolatum.

[0093] Emulsion ointment bases are water-in-oil (W / O) emulsions or oil-in-water (O / W) emulsions and include, for example, acetyl alcohol, stearyl monostearate, lanolin, and stearic acid. Preferred water-soluble ointment bases are prepared from polyethylene glycols of varying molecular weights; see Remington: The Science and Practice of Pharmacy for further information.

[0094] Paste is a semi-solid dosage form in which the active agent is suspended in a suitable base.Depending on the nature of the base, paste is divided into fatty paste or one made from a single-phase aqueous gel.The base in fatty paste is generally petrolatum or hydrophilic petrolatum, etc.Paste made from a single-phase aqueous gel generally contains carboxymethylcellulose, etc. as a base.

[0095] The accelerators are typically lipophilic co-accelerators, also known as "plasticizing" accelerators, i.e., accelerators having a molecular weight in the range of about 150 to 1000 and an aqueous solubility of less than about 1 wt. %, preferably less than about 0.5 wt. %, and most preferably less than about 0.2 wt. The Hildebrand solubility parameter δ of the plasticizing accelerator is in the range of about 2.5 to about 10, preferably in the range of about 5 to about 10. Preferred lipophilic accelerators are fatty esters, fatty alcohols, and fatty ethers. Examples of certain, most preferred, fatty acid esters include methyl laurate, ethyl oleate, propylene glycol monolaurate, propylene glycol dilaurate, glycerol monolaurate, glycerol monooleate, isopropyl n-decanoate, and octyldodecyl myristate. Fatty alcohols include, for example, stearyl alcohol and oleyl alcohol, while fatty ethers include compounds in which a diol or triol, preferably a C2-C4 alkane diol or triol, is substituted with one or two fatty ether substituents. Additional penetration enhancers are known to those skilled in the art of topical drug delivery and / or are described in relevant textbooks and literature. See, for example, Percutaneous Penetration Enhancers, Smith et al. (eds.) (CRC Press, 1995), which is incorporated herein by reference in its entirety.

[0096] In addition to those identified above, various other additives may be included in the compositions of the present invention, including, but not limited to, antioxidants, astringents, fragrances, preservatives, emollients, pigments, dyes, moisturizers, propellants, and sunscreens, as well as other classes of materials whose presence may be pharmaceutically or otherwise desirable. Typical examples of optional additives for inclusion in the formulations of the invention are: preservatives such as sorbates; solvents such as isopropanol and propylene glycol; astringents such as menthol and ethanol; emollients such as polyalkylene methyl glucosides; humectants such as glycerin; emulsifiers such as glycerol stearate, PEG-100 stearate, polyglyceryl-3-hydroxylauryl ether, and polysorbate 60; other polyhydroxy alcohols such as sorbitol and polyethylene glycol; sunscreens such as octyl methoxyl cinnamate (commercially available as Parsol MCX) and butyl methoxybenzoylmethane (available under the trademark Parsol 1789); ascorbic acid (vitamin C), α-tocopherol (vitamin E), β-tocopherol, γ-tocopherol, δ-tocopherol, ε-tocopherol, ζ ι -Tocopherol, Z ΛAntioxidants such as tocopherol, η-tocopherol, and retinol (vitamin A); essential oils, ceramides, essential fatty acids, mineral oils, vegetable oils (e.g., soybean oil, coconut oil, the liquid fraction of shea butter, sunflower oil), animal oils (e.g., perhydrosqualene), synthetic oils, silicone oils or waxes (e.g., cyclomethicone and dimethicone), fluorinated oils (commonly perfluoropolyethers), fatty alcohols (e.g., cetyl alcohol), and waxes (e.g., beeswax, carnauba wax, and paraffin wax); skin feel modifiers; and thickeners and structurants such as swelling clays and cross-linked carboxypolyalkylenes commercially available under the Carbopol trademark. Other additives include beneficial agents such as materials that condition the skin (especially the upper layer of the skin in the stratum corneum), maintain its softness by slowing the loss of its moisture content, and / or protect the skin. Such conditioners and moisturizing agents include, for example, pyrrolidine carboxylic acids and amino acids; organic antimicrobial agents such as 2,4,4'-trichloro-2-hydroxydiphenyl ether (triclosan) and benzoic acid; anti-inflammatory agents such as acetylsalicylic acid and glycyrrhetinic acid; antiseborrheic agents such as retinoic acid; vasodilators such as nicotinic acid; melanogenesis inhibitors such as kojic acid; and mixtures thereof. Further additional active agents include, for example, alpha hydroxy acids, alpha keto acids, polymeric hydroxy acids, moisturizers, collagen, marine extracts, ascorbic acid (vitamin C), α-tocopherol (vitamin E), β-tocopherol, γ-tocopherol, δ-tocopherol, ε-tocopherol, ζ ιAntioxidants include α-tocopherol, ζ2-tocopherol, η-tocopherol, and retinol (vitamin A), and / or pharmaceutically acceptable salts, esters, amides, or other derivatives thereof. A preferred tocopherol compound is α-tocopherol. Additional agents include those that can improve oxygen supply to skin tissue, such as those described in WO 94 / 00098 to Gross et al. and WO 94 / 00109 to Gross et al., both assigned to Lancaster Group AG (incorporated herein in their entirety by reference). Sunscreens and UV-absorbing compounds can also be included. Non-limiting examples of such sunscreens and UV absorbing compounds include aminobenzoic acid (PABA), avobenzone, cinoxate, dioxybenzone, homosalate, menthyl anthranilate, octocrylene, octyl methoxycinnamate, octyl salicylate, oxybenzone, padimate O, phenylbenzimidazole sulfate, sulisobenzone, titanium dioxide, trolamine salicylate, zinc oxide, ensulizole, melazilate, octinoxate, octisalate, and octocrylene. See Title 21, Chapter 1, Subchapter D, Part 352, "Sunscreen drug products for over-the-counter human use," which is incorporated herein in its entirety. Other examples include: Embodiments may include various non-carcinogenic, non-irritating healing materials that facilitate treatment with the formulations of the present invention. Such healing materials may include nutrients, minerals, vitamins, electrolytes, enzymes, herbs, plant extracts, glandular or animal extracts, or safe therapeutic agents that can be added to the formulation to facilitate healing of skin disorders.

[0097] The present invention contemplates amounts of these various additives equivalent to those conventionally used in the cosmetic field, for example, ranging from about 0.01% to about 20% of the total weight of the topical formulation.

[0098] The formulations of the present invention may also contain conventional additives such as opacifiers, flavors, colorants, stabilizers, surfactants, etc. In certain embodiments, other agents such as antimicrobial agents may also be added to prevent spoilage during storage, i.e., to inhibit the growth of microorganisms such as yeasts and molds.

[0099] Suitable antimicrobial agents for the present invention include, but are not limited to, those selected from the group consisting of methyl and propyl esters of p-hydroxybenzoic acid (i.e., methyl and propyl paraben), sodium benzoate, sorbic acid, imidurea, and combinations thereof. In other embodiments, repressors and inducers, i.e., other agents for inhibiting (i.e., glucose) or inducing (i.e., xylose) the production of the polypeptide of interest, can also be added. Such additives can be used provided they are compatible with and do not inhibit the function of the formulation.

[0100] The formulation may also contain mild additives to minimize or eliminate the possibility of skin irritation or damage resulting from the administered chemicals or other components of the composition.

[0101] Suitable irritation-reducing additives include, for example, α-tocopherol; monoamine oxidase inhibitors, particularly phenyl alcohols such as 2-phenyl-1-ethanol; glycerin; salicylate; ascorbate; ionophores such as monensin; amphoteric amines; ammonium chloride; N-acetylcysteine; capsaicin; and chloroquine. When present, the irritation-reducing additive can be included in the composition at a concentration effective to reduce irritation or skin damage, and typically accounts for about 20 wt% or less of the formulation, more typically about 5 wt% or less. Additional suitable pharmacologically active agents that can be included in the formulations in certain embodiments and thus applied topically along with the active agent include, but are not limited to, the following: agents that improve or eradicate pigmented or non-pigmented age spots, keratinocytes, and wrinkles; antimicrobial agents; antibacterial agents; antipruritic and anti-drying agents; anti-inflammatory agents; local anesthetics and analgesics; corticosteroids; retinoids; vitamins; hormones; and antimetabolites.Some examples of topical pharmacologically active agents include acyclovir, amphotericin, chlorhexidine, clotrimazole, ketoconazole, econazole, miconazole, metronidazole, minocycline, nystatin, neomycin, kanamycin, phenytoin, para-aminobenzoic acid esters, octyl methoxycinnamate, octyl salicylate, oxybenzone, dioxybenzone, tocopherol, tocopheryl acetate, selenium sulfide, zinc pyrithione, diphenylhydramine, pramoxine, lidocaine, procaine, erythromycin, tetracycline, clindamycin, crotamiton, hydrochloride, hydroxybenzoates ... These include quinones and their monomethyl and benzyl ethers, naproxen, ibuprofen, cromolyn, retinol, retinyl palmitate, retinyl acetate, coal tar, griseofulvin, estradiol, hydrocortisone, hydrocortisone 21-acetate, hydrocortisone 17-valerate, hydrocortisone 17-butyrate, progesterone, betamethasone valerate, betamethasone dipropionate, triamcinolone acetonide, fluocinonide, clobetasol propionate, minoxidil, dipyridamole, diphenylhydantoin, benzoyl peroxide, and 5-fluorouracil. Creams, lotions, gels, ointments, pastes, etc. can be spread on the affected surface and gently massaged in. Solutions can be applied in the same way, but more typically are applied using a dropper, swab, etc., and carefully applied to the affected area.

[0102] Application regimen depends on several factors that can be easily determined, such as the severity of the condition and its response to initial treatment, but usually will not include application more than twice a day.Those skilled in the art can easily determine the optimal dosage of the formulation to be administered, administration method and repetition rate.Generally, it is contemplated that the formulation of the present invention is applied in the range of once or twice a week to once a day at most.

[0103] III. METHODS AND KITS OF THE INVENTION Treatment method The present invention provides a method for treating a skin disease, comprising administering to a subject in need of such treatment a genetically engineered microorganism, e.g., a genetically engineered bacterium, capable of expressing a recombinant therapeutic fusion protein of the present invention, thereby treating the subject. In a preferred embodiment, the disease is atopic dermatitis. In yet another preferred embodiment, the recombinant therapeutic fusion protein comprises filaggrin. In another embodiment, the recombinant therapeutic fusion protein comprises filaggrin operably linked to a cell-penetrating peptide. In a further embodiment, the recombinant therapeutic fusion protein is operably linked to a translocation signal.

[0104] kit The present invention also provides kits. In one aspect, the kits of the present invention comprise (a) a composition of the present invention and (b) instructions for its use. In another aspect, the kits of the present invention comprise (a) any of the live biotherapeutic compositions of the present invention and (b) instructions for its use. The instructions may include instructions for how to apply, administer, use, and maintain the composition. The compositions of the present invention are as described above. In some embodiments, the compositions of the present invention are engineered microorganisms capable of expressing a therapeutically relevant recombinant fusion polypeptide, as described above. In a preferred embodiment, the composition comprises an engineered bacterium (e.g., Staphylococcus epidermidis) capable of expressing a recombinant fusion polypeptide comprising filaggrin.

[0105] In some embodiments, the kit may include a sealed container. Non-limiting examples of containers include bottles, metal tubes, laminated tubes, plastic tubes, dispensers, pressurized containers, barrier containers, packages, compartments, lipstick containers, compact containers, cosmetic trays capable of holding cosmetic compositions, or other types of containers such as syringe bottles or blow-molded plastic containers in which dispersions or compositions or desired bottles, dispensers, or packages are held. Other examples of containers include glass or plastic vials or bottles. The kit and / or container may include indicia on its surface. The indicia may be, for example, a word, phrase, abbreviation, picture, or symbol.

[0106] The present invention is further illustrated by the following examples, which should not be construed as further limiting. All figures and the contents of all references, patents and published patent applications, and figures cited throughout this application are expressly incorporated herein by reference in their entirety. (Example)

[0107] The following examples further describe and demonstrate embodiments within the scope of the present invention. The examples are given for illustrative purposes only and should not be construed as limitations of the invention, as many variations thereof are possible without departing from the spirit and scope of the invention. [Example]

[0108] Development of nucleic acid constructs capable of encoding proteins that can be exported from Staphylococcus epidermidis cells and then imported into human keratinocytes In one embodiment, the present invention describes the generation of recombinant S. epidermidis strains that can secrete heterologous proteins, thus overcoming the recalcitrance of genetic modification of S. epidermidis. Functional genetic analysis of S. aureus and S. epidermidis, common skin colonizers, has previously been limited due to the presence of Type I and Type IV restriction systems in virtually all strains of these bacteria. These restriction systems recognize methylated cytosine bases in DNA derived from standard clonal expansion systems, such as DH10B E. coli. However, using a methylation-deficient E. coli strain, S. epidermidis strain ATCC12228, a commensal, non-pathogenic isolate lacking the ica operon implicated in S. epidermidis-associated catheter bloodstream infections, was successfully performed. 5 Several constructs were created in the study. Thus, the present invention describes the first known reported heterologous protein expression in S. epidermidis. The present invention thus provides, in one embodiment, a nucleic acid plasmid capable of encoding a protein that is exported from S. epidermidis cells and subsequently imported into human keratinocytes. This plasmid, pAZT, is based on the allelic exchange E. coli-Staphylococcus shuttle vector pJB38 (Bose, JL et al., Applied and environmental microbiology. 2013;79(7):2218-2224), which has been specifically re-engineered to have features that improve its function. The present invention, in one embodiment, provides a plasmid capable of effectively colonizing the reconstituted epidermis and capable of expressing a heterologous protein in a cell culture medium that can express ... 9 We provide engineered S. epidermidis strains capable of producing 50 μg of protein per mL of CFU / mL (Figure 2).

[0109] Cell-penetrating peptides (CPPs) Despite the formidable barrier properties of the epidermis, protein delivery through the stratum corneum has been demonstrated using transduction or cell-penetrating peptides (CPPs). This challenge arises due to the hydrophobic surface and impaired diffusion from the connected corneocyte layers, including the stratum corneum. However, by attaching a CPP sequence to the N-terminus of a protein of interest, successful delivery of the fusion protein to deeper epidermis is possible, in addition to facilitating the target protein's intracellular localization and endosomal / lysosomal escape. In one embodiment, the present invention provides a construct using such an approach, including a transcription-derived cell-penetrating peptide (RMR) protein motif HIV transactivator (Figure 2). This approach provides the basis for delivering proteins to deeper layers of the skin for enhanced therapeutic efficacy.

[0110] Safety and "Kill Switch" A key requirement for nearly all recombinant microorganisms for clinical use is the ability to prevent unwanted introduction into other individuals or environments. To ensure the safety of the engineered strains, in one embodiment, the present invention uses auxotrophic strains that require supplementation of critical amino acids (D-ala) or certain metabolic genes (AlaR) for survival, simultaneously replacing the need for antibiotic resistance strains for selection, although the latter is not commercially viable. In another embodiment, the present invention incorporates a "kill switch" based on CRISPR / Cas9 self-cleavage upon induction of a dual xylose-riboswitch promoter. In yet another embodiment, the present invention provides a cell counter that recombines from the AZT locus after a defined number of divisions, but this method requires reapplication of the vehicle. To ensure the safety of the engineered S. epidermidis of the present invention, we developed a CRISPR / Cas9-based kill switch that is xylose-inducible and dual-regulated using a theophylline riboswitch. The basis of this approach is that Cas9 is extremely efficient at chromosomal cleavage given a targeted guide, and because Staphylococcus aureus lacks the standard non-homologous end joining repair pathway, genome cleavage leads to cell death in the absence of a homologous recombination template. The use of a CRISPR-based system also provides high specificity, since comparative genomics can be used to design unique guides for the engineered Staphylococcus epidermidis strain of the present invention, so that the construct is inactive when spread to other microorganisms by horizontal gene transfer. Finally, in one embodiment, the present invention provides a construct designed to express multiple CRISPR spacers to simultaneously target and reliably cleave multiple genomic regions and minimize survival by reversion. [Example]

[0111] Determining the persistence and localization of topically applied Staphylococcus epidermidis using an in vitro model system Materials & Methods Creation of reporter bacteria To facilitate tracking of topically applied bacteria, we used an sGFP-expressing strain of Staphylococcus epidermidis (SE). The SecA and RMR peptides were removed to prevent the sGFP protein from being shuttled into the secretory system and to prevent free sGFP from penetrating the stratum corneum. This construct is referred to as SE-sGFP.

[0112] Quantification and comparison of growth characteristics of transformed bacteria in liquid media A basic understanding of the ability of transformed (recombinant) S. epidermidis to compete with wild-type S. epidermidis was required. To understand the growth characteristics of the transformed bacteria and the growth kinetics of the recombinant, protein-producing bacteria, colony-forming units (CFU) were quantified in liquid medium using standard techniques. To determine the growth differences between S. epidermidis-sGFP, S. epidermidis-chl, and wild-type S. epidermidis, each strain was grown separately in two triplicate 100 mL cultures for 12 h each. Every hour, 1 mL samples were taken and measured at 395 nm and 600 nm to obtain both sGFP signal and total bacterial concentration measurements, respectively. Fluorescence and optical density were compared across all samples to understand growth characteristics and sGFP production. Results indicated that protein production only slightly reduced competitive growth of S. epidermidis-GFP compared to S. epidermidis-chl, as determined by fluorescence and CFU measurements.

[0113] Quantification of growth of S. epidermidis-GFP and control strains on RHE To characterize the feasibility of applying bacteria to the skin, we determined the growth kinetics of externally applied bacteria on an in vitro skin model, understanding that this is only a first approximation of the ecological competition these bacteria encounter on human skin. The assay began two days after receiving differentiated cultures to allow the cultures to stabilize after shipping. RHE cultures were established and maintained in antibiotic- and antimycotic-free medium (supplemented with Chl as needed), which was changed every two days. Bacteria suspended in 50% glycerol were applied to the central 3 mm diameter of the RHE using a pipette. Control RHE with S. epidermidis-Chl and S. epidermidis-WT bacteria were also applied and removed simultaneously with the experimental arm. Upon removal from culture, the tissue insert was homogenized and passed through a 5 μm filter to allow collection of bacterial effluent. The bacterial suspension was spun down, resuspended in medium, serially diluted, and plated to determine the CFU of bacteria in the insert. All measurements were normalized by the maximum bacterial recovery determined by the CFU recovered 15 minutes after application.

[0114] Qualitative characterization of growth of SE-GFP and control strains on RHE An assay was designed to obtain spatial and temporal information about S. epidermidis-GFP using RHE and a Vivascope. S. epidermidis-GFP was applied to the RHE, and the sample was imaged in reflectance and fluorescence modes over three standardized regions, 2 mm x 2 mm wide and 100 μm deep, using 10 μm steps and a linear increase in laser power. Ultrasound gel (Parker Laboratories) was used to maintain the refractive index between the object and the glass sample plate. Images were analyzed in ImageJ using "Grid / Collection Stitching" (Figure 3).

[0115] The results showed that the bacteria progressed towards the surface and deep grooves of the stratum corneum and remained constant over the course of the experiment.

[0116] Importantly, to mimic the ultrastructure of damaged skin in patients with atopic dermatitis, RHE was intentionally punctured with a Derma Microneedle device to determine bacterial localization in the presence of damaged skin. Results showed that bacteria localized to the puncture site at a depth of up to 70 µm (arrows, Figure 3(B)-(D)). This suggests that topically applied bacteria can travel toward areas of damaged skin.

[0117] These studies were repeated in vivo. Specifically, SE-GFP was applied and observed by optical and in vivo two-photon microscopy 3 days after application. The results showed persistent and widespread GFP expression at depths ranging from 25 μm in mouse ears to 80 μm in the shaved dorsal skin of mice (Figure 4). This demonstrates the ability of S. epidermidis-GFP to colonize the deepest layers of the stratum corneum (10–40 μm) and further colonize mouse hair follicles. [Example]

[0118] Characterization of bacterially secreted sGFP delivery to skin using an in vitro model system Characterization of sGFP production in SEs Characterization of delivery of bulk-purified sGFP and sGFP+RMR to the RHE Data on the localization of purified sGFP and sGFP+RMR will facilitate understanding (i) whether sGFP+RMR penetrates the stratum corneum; (ii) if so, how deeply it can be detected; and (iii) the dynamics of penetration. Here, 5.0 μg / μL of GFP+ / -RMR was applied at 0, 2, 6, 12, 18, and 24 hours to determine the effects of dose on penetration and time. Results showed that GFP was detected deep at the epidermal-dermal junction within 30 minutes of application.

[0119] Characterization of cellular compartments and penetration depth of in situ secreted sGFP proteins from SE-GFP reporter and control strains The goal of this assay is to identify the protein SE-sGFP. RMR / SecA and SE-sGFP SecA This method is similar to that described above, with the important difference that the cells were generated in situ by IHC and compared to a control strain on RHE. Using the same method described herein, we characterized the kinetics of sGFP penetration into RHE. While the Vivascope provided useful information about sGFP penetration across a larger surface area, the use of IHC allowed for more subtle discrimination. The IHC method can detect GFP-positive regions relative to peptidoglycan-positive regions in the SE, allowing secreted sGFP to be distinguished from the sGFP signal in bacteria (Figure 5).

[0120] Non-fluorescent detection of therapeutic proteins. The therapeutic protein ultimately delivered may not be fluorescent, and suitable antibodies may not be available to characterize its delivery to the epidermis. Proteomic analysis of stratum corneum tape strips was used to characterize differences in in vivo protein profiles from patients with atopic dermatitis (Sakabe, J. et al., The Journal of allergy and clinical immunology. 2014;134(4):957-960 e958) and ichthyosis (Rice, R. H. et al., PloS one. 2013;8(10):e75355). Furthermore, horizontal sections from the inside to the outside of skin biopsies can be used to demonstrate the ability to identify low-abundance molecules after penetration into the stratum corneum. [Example]

[0121] Evaluation of the pharmacokinetics (PK) and pharmacodynamics (PD) of AZT-01 in mice (non-GLP) The PK / PD of AZT-01 was evaluated using a genetic atopic dermatitis mouse construct (flaky tail mice), whereas the PK of AZT-01 was evaluated in healthy mice. To assess PD, AZT-01 was topically applied to mice, and PD was assessed by phenotype (erythema, edema, exfoliation, dryness, and transepidermal water loss) and histological changes (skin barrier remodeling, fibrosis, CD4+ T cells, etc.). To assess PK, the distribution of AZT-01 and filaggrin was examined, and colonization patterns were characterized. Changes in the skin microbiota were evaluated.

[0122] Atopic Dermatitis Mouse Model. To investigate the applicability of the method, two model mouse strains were used: filaggrin knockout mice (flg- / -) and flaky tail mice (ft / ft). A complete review of atopic dermatitis mouse models is presented by Geha et al. (Vavrova K. et al., The Journal of Invest. Dermat. 2014;134(3):746-753). Briefly, flg- / - mice exhibit dry, scaly skin. Despite a significant reduction in natural moisturizing components, which are filaggrin degradation products, the moisture content of the stratum corneum (SC) and transepidermal water loss (TEWL) were normal in flg- / - mice. Antigens penetrated flg- / - SC more efficiently, resulting in enhanced responses in hapten-induced contact hypersensitivity and higher serum levels of anti-ovalbumin IgG(I) and IgE. Therefore, mouse ears were sensitized with OVA antigen.

[0123] ft / ft mice have two distinct autosomal recessive mutations: a matted hair (ma) and a scaly tail (ft). These mice spontaneously develop dermatitis and exhibit elevated serum IgE levels, even under specific pathogen-free conditions. Flaky tail mice also have a loss-of-function mutation in filaggrin (Flg) and exhibit abnormal skin barrier function, as well as increased TEWL and SC moisture.

[0124] Study design. The study was conducted over a 4-week period with four arms in both types of mice (Flg- / - and ft / ft). Mice were divided into the following treatment groups: topical vehicle control (50% glycerol, 50% sterile BHI medium), topical recombinant filaggrin (purified recombinant filaggrin at 50 μg / ml), topical wild-type Staphylococcus epidermidis (SE) (1.0 x 10 in 50% glycerol ... 9 CFU), and local SE FLG (1.0x10 in 50% glycerol 9 The mice were randomized to 100% (CFU). Each solution was applied to the same ear of each mouse on days 0, 7, 14, and 21, and the mice were also evaluated on these days before application of the appropriate solution. The final evaluation was on day 28, after which time the mice were sacrificed according to the appropriate animal protocol. The primary outcome (described in detail below) was the change in clinical disease score, which assessed gross changes in symptoms.

[0125] Based on a sample size estimate using a standard deviation of 2.5, 90% power, and a type I error of 0.05, 8 mice / arm / genotype were required to detect a mean change in clinical disease score of 4 points between groups, meaning a total of 32 mice per genotype (64 mice total) were required for the study.

[0126] [Table 2]

[0127] Primary outcome: Clinical disease score. SE FLGTo measure the effects of treatment on macroscopic and microscopic changes associated with ear treatment, a clinical score based on previous studies was used (see, e.g., Matsuoka H. et al., Allergy. 2003;58(2):139-145, and Kim, MC et al., Journal of acupuncture and meridian studies. 2013;6(2):98-109). The composite clinical disease score was the sum of the severity of erythema, edema, peeling, and dryness on the ear surface and was scored accordingly as 0 (not visible), 1 (mild), 2 (moderate), and 3 (severe). Scoring was performed by individuals who were blinded to the treatment status of each group. The skin was photographed every 7 days. Additionally, TEWL was measured using a TEWL meter (Khazaka Electronic). Finally, we evaluated its ability to disrupt the skin barrier and prevent the penetration of foreign substances through the stratum corneum (SC) using a method described by Kawasaki (i.e., Kawasaki, H. et al., The Journal of Allergy and Clinical Immunology. 2012;129(6):1538-1546.e1536). Calcein bis[N,N-bis(carboxymethyl)aminomethyl]fluorescein (Sigma-Aldrich) was mixed with liposomes prepared from Presome CSII-101 (Nippon Fine Chemical, Osaka, Japan) and applied topically to the cutaneous area of ​​6- to 8-week-old mice for 3 hours. The tails were then removed and rapidly frozen and embedded.

[0128] Characterization of the microbiota after AZT-01 application. To understand the impact of SE addition on the microbial diversity of the skin microbiota, we measured changes in the microbial community using combined 16S rRNA analysis. This was performed at the Jackson Laboratory's JAX Genomic Medicine Facility using the Illumina MiSeq platform with RT-PCR and qPCR for sequencing. Changes in the relative abundance of bacteria in the microbial community were measured using the method described by Caporaso et al. (Caporaso, JG et al., The ISME journal. 2012;6(8):1621-1624). Briefly, samples were collected from the skin using cotton swabs, and rRNA was extracted using an rRNA extraction kit (Qiagen), then amplified, analyzed using qPCR, and sequenced. Similar sequences were then grouped into operational taxonomic units (OTUs) using bioinformatics and statistical methods (Figure 6).

[0129] Ecological metrics and community structure analysis were used to measure dysbiosis. First, dysbiosis was assessed as a function of diversity using the Shannon Diversity Index, an ecological measure of the microbial community under consideration, and compared before and after application. Furthermore, the community structure of the local microbiota was compared before and after treatment. Then, dysbiosis was measured as (i) the overall deviation (%) from the baseline microbiota, and (ii) the deviation from the average community structure across our controls using statistics such as the Yue-Clayton index to compare community structure. Finally, microbiota trends were analyzed at the per-species level. The long-term dynamics of each species was also tracked across treatments to identify whether species were lost from the community.

[0130] Immunohistochemistry study. Filaggrin was visualized and quantified using immunohistochemistry by comparing AZT-01 with the vehicle control. Keratinocyte cells were fixed with 70% ethanol and 50 mM glycine for 1 hour. Immunofluorescence staining was performed by incubating with anti-filaggrin primary antibody at 1:200 dilution for 2 hours, followed by incubation with rat anti-goat rhodamine secondary antibody (Jackson Laboratory) at 1:200 dilution in the presence of Hoechst Stain Solution (Sigma). Slides were coverslipped in Gel / Mount (Biomed). Additionally, alternative sequences were created and tested in place of RMR signals (e.g., endosomal escape peptides such as those described in Appelbaum et al., 2012, the entire contents of which are incorporated herein by reference) (Appelbaum, JS et al., Chemistry & Biology. 2012;19(7):819-830).

[0131] Statistical Analysis. Differences between groups in the primary outcome and / or gross clinical disease scores were assessed using a two-tailed Student's T-test. Differences between groups were assessed using ANOVA. The same technique was used to assess TEWL and epidermal thickening. Differences in nonparametric continuous variables were assessed using the Mann-Whitney U test. Finally, differences in ordinal variables were assessed using the chi-square test.

[0132] For microbiota analysis, community variation between samples was calculated using quantitative, taxonomic-based Canberra distance. Discriminant analysis of intragroup similarity was performed using permutation MANOVA. To determine whether skin microbial communities become more similar to each other, we used a β-variance test using the betadisper function in vegans. This test is a multivariate analog of Levene's test of homogeneity of variance and tests for significant differences in sample heterogeneity between groups (Anderson, MJ et al., Ecology Letters. 2006;9(6):683-693). P values ​​for significant indices were adjusted for multiple comparisons using Holm's correction (Holm S. Scandinavian Journal of Statistics. 1979;6(2):65-70).

[0133] Immunofluorescence microscopy. Filaggrin localization was visualized using immunofluorescence microscopy. Mouse skin samples were fixed in 10% formalin and embedded in paraffin. Paraffin sections were dewaxed and washed with 95% ethanol, followed by methanolic hydrogen peroxide. Sections were then treated with a heat-induced epitope retrieval (HIER) procedure using rodent Decloaker solution (Biocare Medical, RD913) and a Biocare decloaking chamber. After washing in Tris pH 7.4, sections were incubated with rat serum and FcBlock (24G2) diluted in blocking solution, followed by rabbit anti-E. coli B (DAKO, B0357). Samples were washed in Tris and then incubated with goat anti-rabbit IgG-Texas Red antibody (Invitrogen, T2767). Tissues were then counterstained with HOECH™ and imaged using a Leica DM IRBE fluorescence microscope. [Example]

[0134] AZT-01 and its effects on local and systemic inflammation and immunity in rats (non-GLP) To assess toxicity, local tissue samples, serum samples, and lymph node samples were analyzed from topically treated rats euthanized at specific time points. Tissues were analyzed for histological changes in inflammatory activity (e.g., levels of CD4+ T cells, Langerhans cells, IgE, IL-4, IFN-γ, etc.) and activity and changes in skin immune responses (e.g., levels of IL-4, IL-10, IL-13, etc.). Clinical signs of potential side effects associated with the use of therapy, including erythema, skin temperature changes, edema, blistering, and ulceration, were also monitored.

[0135] Histological evaluation. The excised ears from each group were fixed in 4% paraformaldehyde for 16 h and embedded in paraffin. Subsequently, 6 μm sections were stained with hematoxylin (Sigma Aldrich, St. Louis, MO, USA) and eosin (Sigma Aldrich, St. Louis, MO, USA) (H&E). Lymphocyte infiltration and fibrosis in the dermis were observed under a microscope (100X, 200X).

[0136] Quantification of disease-related mRNA transcripts. Changes in the expression of genes associated with the onset, progression, and maintenance of atopic dermatitis (AD-associated pathogenic cytokines (e.g., IL-4, IL-5, IL-13, IFN-γ, IL-17, IL-10, and TNF-α)) were measured by standard qPCR assays. Briefly, total RNA from skin samples was isolated using the Qiagen RNeasy Mini Kit (Qiagen, Valencia, CA) according to the manufacturer's instructions. Respective cDNAs were synthesized using reverse transcriptase PCR (RT-PCR). Real-time PCR was performed using the comparative 2-ΔΔCT method and normalized to glyceraldehyde 3-phosphate dehydrogenase (GAPDH) transcript levels.

[0137] Immunological changes after AZT-01 application. Cells from the bacterial- and non-bacterial-treated areas were isolated to examine immunological changes. Keratinocytes, epidermal cells, lymph node-derived cells, and small intestinal lamina propria were isolated. Single-cell suspensions were stained with the LIVE / DEAD Fixable Blue Dead Cell Stain Kit (Invitrogen) or 4',6-diamidino-2-phenylindole (DAPI, Sigma) in HBSS to remove dead cells. For transcription factor detection, cells were stained using the Foxp3 staining set (eBioscience) according to the manufacturer's protocol. For intracellular cytokine detection, cells were fixed, permeabilized with BD Cytofix / Cytoperm, and stained in BD Perm Wash buffer (BD Biosciences). Cells were stained with the following antibodies purchased from eBioscience, BD Biosciences, or Dendritics Corp: CD4, IL-10, IL-17A, IFN-γ, TNF-α, Foxp3, CD34, CD44, and / or CD25. Staining was performed in the presence of FcBlock (eBioscience), 0.2 mg / ml purified rat IgG, and 1 mg / ml normal mouse serum (Jackson Immunoresearch). Staining for skin homing markers was performed as previously described (Lopes, LB et al., Pharm Res. 2005;22(5):750-757). As a measure of safety, signs of hematogenous AZA-01 were monitored by collecting samples of the skin-draining lymph nodes (DLN) and spleen of one mouse two weeks after application of AZT-01. Briefly, mice were placed in 70% EtOH and moved to a laminar flow hood. After 1–2 min in EtOH, the DLN and spleen were isolated in a 50 mL conical tube with a 70 μm strainer and processed as separate samples. The organs were dissociated in 500 μl of sterile PBS, and 50–100 μl of the cell suspension was plated on BH agar. [Example]

[0138] Performing initial formulation and analytical method development to evaluate proposed specification sets for active pharmaceutical ingredients (API) and drug products (DP) Critical to the clinical development of live biotherapeutic products (LBPs; e.g., AZT-01) is developing useful formulations and practical analytical assays that consider the unique properties of LBPs as active pharmaceutical ingredients (APIs) or drug products (DPs) compared to traditional small molecules. Develop formulations for the API, manufacture the DP, develop analytical methods for both the API and DP, and establish GMP material specifications for clinical trials.

[0139] Statistical analysis. Unless otherwise indicated, experiments were performed in triplicate, and means and standard deviations are reported. For comparisons between groups, two-tailed t-tests or analysis of variance were used. When data were not normally distributed, they were replaced with non-parametric equivalents (Wilcoxon rank sum and Kruskal-Wallis test). [Example]

[0140] Hydrophobicity analysis Human filaggrin (hFLG) contains 12 repeating units and is processed by intracellular proteases to release individual units. The sequences within these units are highly homologous. It has been proposed that the units are cleaved at "linker regions." The segments between these linker regions are individual units, each of which contains a pair of subdomains. The entire human filaggrin (hFLG) sequence (Uniprot P20930) was analyzed using the online Kite-Doolittle calculation tool (https: / / web.expasy.org / protscale / ). The hydrophobicity score as a function of amino acid position graph is shown in Figure 7. As shown in Figure 7, there is a periodic increase in hydrophobicity. The Kite-Doolittle score gave the highest difference between the highest and lowest values. When compared to the mouse filaggrin (mFLG) sequence (NCBI Reference Sequence: XP 017175331.1) shown in Figure 8, a much higher difference in hydrophobicity score is observed. This is due to the nature of proteins from different organisms.

[0141] Further examination of the molecular regions of interest revealed that the distance between vertices corresponding to more hydrophobic regions was greater in human filaggrin than in mouse, and the maximum hydrophobicity was higher in mouse than in human.

[0142] Two specific domains of hFLG were of interest: hFLG [domains 5-6] and [domains 9-10]. Figure 9 shows the region of hFLG starting at amino acid 1400 to 1800, showing the unit segments of domains 9 and 10. The repeat ends of the peaks immediately following the 10-amino acid segment (SGSQASDSEGHS) common to human and mouse filaggrin are also shown. The high peaks at positions 1444 and 1767 on the graph correspond to the FLY segment. The peaks at 1679 and 1929 are regions containing polytyrosine.

[0143] The hFLG protein (domains 9-10) (1429-1774) (SEQ ID NO: 2) represents a highly hydrophobic segment. This segment is shown in Figure 10. It contains FLY segments at both ends of the protein.

[0144] SEQ ID NO: 2 JPEG2024009822000005.jpg92142

[0145] The complete sequence of the hFLG protein (domains 9-10) (1429-1774) with an N-terminal methionine and an RMR segment at the C-terminus is shown in SEQ ID NO: 3 (M and RMR are underlined). The repeat sequence QSGEXSGRSXSFLYQVSXHEQSES is shown below in bold. SEQ ID NO: 3 JPEG2024009822000006.jpg72144JPEG2024009822000007.jpg20150

[0146] It was found that the repetition of the FLY segment is not necessary for activity and, furthermore, it is a burden to the bacterium and may be the basis for the lack of production in Gram-positive organisms.

[0147] FIG. 10 is a graph showing the hydrophobicity score as a function of amino acid position for hFLG[9-10] (1429-1774) from amino acid position 1400 to amino acid position 1800 in hFLG. [Example]

[0148] New protein structures based on hydrophobicity analysis hFLG [domains 9-10] Next, we also produced hFLG [domains 9-10] proteins in which the N-terminal region was systematically truncated.

[0149] Selecting segments based on a "U" ("high-high") shape The graph in Figure 11 shows that the start and end positions of hFLG[9-10](1429-1777) (SEQ ID NO: 2) may be too long. Therefore, the N-terminal FLY segment was truncated to generate hFLG[9-10](1452-1777) (SEQ ID NO: 4) to prevent undesired recombination events. SEQ ID NO:4 JPEG2024009822000008.jpg77142 Theoretical pI / Mw: 10.65 / 36162.11

[0150] Selecting segments based on "A" ("low-high-low") shape Next, a selection of repeat sequences from a point of low hydrophobicity to the next point of low hydrophobicity was performed. This segment has two domains at the N-terminus with the FLY region near the middle of the sequence. Thus, the "core" sequence containing domains 9-10 is at the end of the sequence rather than the middle. Furthermore, the long cross-repeat sequence is toward the C-terminus of the sequence. This is shown in Figure 12.

[0151] Based on the above analysis, a protein based on the low-low segment hFLG[9-10](1545-1869) was generated and is shown below as SEQ ID NO:5. SEQ ID NO:5 JPEG2024009822000009.jpg73144Theoretical pI / Mw: 11.00 / 36182.19

[0152] The importance of the segment with no known function was assessed by removing it in hFLG[9-10](1545-1777), shown below as SEQ ID NO:6. SEQ ID NO:6 JPEG2024009822000010.jpg53144 Theoretical pI / Mw: 10.07 / 26329.02

[0153] It is further contemplated to knock off the N-terminal region on hFLG[9-10](1452-1777) (SEQ ID NO: 2). [Example]

[0154] Filaggrin (M-AZT-mutant protein-RMR) The hFLG[9-10] sequence (SEQ ID NO: 1) was subjected to BLAST analysis: Filaggrin [human] Sequence number: NP_002007.1 Length: 4061, Matches: 16 Range 1:1430-1774. JPEG2024009822000011.jpg208142

[0155] This sequence was compared with other hFLG units (FLG[3-4], hFLG[5-6], hFLG[7-8], hFLG[11-12], hFLG[13-14], hFLG[15-16], hFLG[17-18], hFLG[19-20], and hFLG[21-22]). Amino acids that were variable were compared, and the most common ones were replaced in the [9-10] sequence.

[0156] The new sequence (S-FLG) was compared with the original hFLG [9-10] by multiple alignment and is shown below: JPEG2024009822000012.jpg199115hFLG[M-AZT-mutate-RMR](1-352) was created and is shown below as SEQ ID NO:8.

[0157] JPEG2024009822000013.jpg76146

[0158] The amino acids shown in bold in SEQ ID NO: 8 were modified to the most frequently occurring amino acids when all FLG units were compared. Table 3 below shows the amino acid residues in hFLG[9-10] and the corresponding modified amino acids in hFLG[AZT-mutant protein]. Ser-Glu-Ser (SES) was reintroduced into the sequence and aligned using Kite-Doolittle analysis. For the same reason as SES, the QSGESSG sequence was removed and calculated to be unfounded.

[0159] [Table 3] [Example]

[0160] Filaggrin consensus sequence A human filaggrin consensus sequence was generated by alignment of the filaggrin dimers hFLG[3-4], hFLG[5-6], hFLG[7-8], hFLG[9-10], hFLG[11-12], hFLG[13-14], hFLG[15-16], hFLG[17-18], hFLG[19-20], and hFLG[21-22], as shown in Figure 13. The consensus sequence shown as SEQ ID NO: 9 refers to the sequence formed from the most frequently occurring amino acids shown in the alignment in Figure 13. SEQ ID NO:9 JPEG2024009822000015.jpg43163 [Example]

[0161] Keratin-binding assay to measure filaggrin activity The activity of various hFLG[9-10] sequences set forth in SEQ ID NOs: 1-9 was measured using a keratin binding assay.

[0162] Keratin extraction from human calluses: Keratin was extracted from human calluses. One to two grams of callus was homogenized in 10 mL of 5 mM Tris pH 7.4 containing protease inhibitors using a Bullet Blender bead beater. The homogenized tissue was centrifuged at 10,000 rpm for 20 minutes at 4°C. The resulting pellet was resuspended in 0.05 M Tris pH 7.4 containing 8 M urea and 0.025 mM β-mercaptoethanol and gently mixed at 37°C for 2 hours. The urea lysate was centrifuged at 10,000 rpm for 10 minutes. The supernatant was dialyzed against 5 mM Tris pH 7.4 overnight at 4°C to allow for the gradual assembly of keratin filaments.

[0163] Filaggrin-keratin binding assay: Aliquots of assembled callus keratin filaments (see above section) were centrifuged at 10,000 RPM for 5 minutes to remove insoluble contaminants. A specific amount of keratin extract diluted in coating buffer to a final volume of 200 μL was used to coat wells overnight at 4°C. The coating buffer was removed, and wells were blocked with 200 μL of 5% nonfat dry milk in PBST (PBS + 0.05% Tween 20) and incubated for 2 h at 37°C. The blocking buffer was removed, and a predetermined amount (μg) of the appropriate human recombinant filaggrin protein diluted in PBST to a final volume of 200 μL was added, and the plate was incubated for 2 h at 37°C. The plate was washed three times with 200 μL of PBST per well. For detection of filaggrin binding, an anti-filaggrin IgY chicken antibody (RL-012-001B diluted 1 / 1000 in PBST) was added to the wells. For detection of keratin (control), 200 μL of mouse pan-keratin antibody (type II AE3 Ab diluted 1 / 500 in PBST) was added to the indicated wells. The primary antibody was incubated for 1 h at room temperature with shaking. The primary antibody was removed, and the plate was washed three times with PBST. A secondary alpaca anti-chicken antibody conjugated to horseradish peroxidase (HRP) was added at a 1 / 2000 dilution in PBST to all wells containing the anti-chicken antibody. Wells containing the anti-keratin antibody received a goat anti-mouse antibody conjugated to HRP diluted 1 / 500 in PBST. The plate was incubated for 1 h at room temperature with shaking. The plate was washed three times with PBST, and 100 μL of 1-Sterp Ultra TMB substrate was added. The plate was incubated at room temperature for 15–20 min until the appearance of the blue color subsided. The reaction was stopped by adding 100 μL of 2 M HCl and the absorbance was read on a spectrophotometer at 450-550 nm.

[0164] The results, shown in Figures 14-16, demonstrate differences in binding to keratin among the hFLG sequences tested. Figure 14 is a graph showing background FLG binding at 1 μg / well over 2 hours at 37°C. The results shown in Figure 14 include nonspecific binding (NSB). Figure 15 is a graph showing binding of various hFLG segments to human callus keratin (NSB removed). Figure 16 is a graph showing titration of IgY anti-hFLG. As shown in Figures 14-16, hFLG[5-6] did not bind to keratin, while the various hFLGs tested [9-10] showed binding to keratin. [Example]

[0165] Activity of SE secreting hFLG[9-10] in mice The colonization kinetics of FLG-producing SE in vivo was evaluated using a genetic IV mouse model (Flg- / -) and wild-type mice. Flg- / - mice are filaggrin-deficient and exhibit dry, scaly skin. Despite a significant reduction in natural moisturizing components, which are filaggrin degradation products, the moisture content of the stratum corneum (SC) and TEWL are normal in Flg- / - mice. Antigens penetrate Flg- / - SC more efficiently, resulting in enhanced responses in hapten-induced contact hypersensitivity and higher serum levels of anti-ovalbumin (OVA) IgG(1) and IgE. Flg- / - mice were obtained from the RIKEN BioResource Research Center (RIKEN BRC, Tsukuba, Ibaraki, Japan). Wild-type mice (BALB / c) were also used in this experiment.

[0166] hFLG[9-10] sequences, set forth in SEQ ID NOS: 1-9, are used in SE in Flg- / - and BALB / c mice. Five groups of each mouse type are used in the study over a 4-week period. Mice are divided into the following treatment groups: topical vehicle control (50% glycerol, 50% sterile BHI medium), topical wild-type SE (1.0x10 in 50% glycerol), and topical wild-type SE (1.0x10 in 50% glycerol). 8 CFU / cm2 ), and three doses of each of the filaggrin-secreting SE constructs (SEFLG) (10 in 50% glycerol) 6 , 10 7 , and 10 8 CFU / cm 2 Each solution is applied to the same ear and tail of each mouse daily for 7 days, and the mice are evaluated for microbiota analysis on days 7, 14, 30, and 60 to assess colonization kinetics, and for microscopy and histology on days 7 and 14 to assess localization in the skin and gross changes (e.g., any signs of adverse events such as inflammation). Twelve mice are used in each arm per mouse type.

[0167] equivalent Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein which equivalents are intended to be encompassed by the following claims.

[0168] The present disclosure includes the following sequence information: SEQUENCE LISTING <110> AZITRA INC <120> METHODS AND COMPOSITIONS FOR TREATING INFLAMMATORY SKIN DISEASE WITH RECOMBINANT MICROORGANISMS <130> PA23-485 <140> <141> 2018-09-05 <150> US 62 / 685,687 <151> 2018-06-15 <150> US 62 / 554,271 <151> 2017-09-05 <160> 52 <170> PatentIn version 3.5 <210> 1 <211> 4061 <212> PRT <213> Homo sapiens <400> 1 Met Ser Thr Leu Leu Glu Asn Ile Phe Ala Ile Ile Asn Leu Phe Lys 1 5 10 15 Gln Tyr Ser Lys Lys Asp Lys Asn Thr Asp Thr Leu Ser Ser Lys Glu 20 25 30 Leu Lys Glu Leu Glu Lys Glu Phe Arg Gln Ile Leu Lys Asn Pro 35 40 45 Asp Asp Pro Asp Met Val Asp Val Phe Met Asp His Leu Asp Ile Asp 50 55 60 His Asn Lys Lys Ile Asp Phe Thr Glu Phe Leu Leu Met Val Phe Lys 65 70 75 80 Leu Ala Gln Ala Tyr Glu Ser Thr Arg Lys Glu Asn Leu Pro Ile 85 90 95 Ser Gly His Lys His Arg Lys His Ser His His Asp Lys His Glu Asp 100 105 110 Asn Lys Gln Glu Glu Asn Lys Glu Asn Arg Lys Arg Pro Ser Ser Leu 115 120 125 Glu Arg Arg Asn Asn Arg Lys Gly Asn Lys Gly Arg Ser Lys Ser Pro 130 135 140 Arg Glu Thr Gly Gly Lys Arg His Glu Ser Ser Ser Glu Lys Lys Glu 145 150 155 160 Arg Lys Gly Tyr Ser Pro Thr His Arg Glu Glu Glu Tyr Gly Lys Asn 165 170 175 His His Asn Ser Ser Lys Lys Glu Lys Asn Lys Thr Glu Asn Thr Arg 180 185 190 Leu Gly Asp Asn Arg Lys Arg Leu Ser Glu Arg Leu Glu Glu Lys Glu 195 200 205 Asp Asn Glu Glu Gly Val Tyr Asp Tyr Glu Asn Thr Gly Arg Met Thr 210 215 220 Gln Lys Trp Ile Gln Ser Gly His Ile Ala Thr Tyr Tyr Thr Ile Gln 225 230 235 240 Asp Glu Ala Tyr Asp Thr Thr Asp Ser Leu Leu Glu Glu Asn Lys Ile 245 250 255 Tyr Glu Arg Ser Arg Ser Ser Asp Gly Lys Ser Ser Ser Gln Val Asn 260 265 270 Arg Ser Arg His Glu Asn Thr Ser Gln Val Pro Leu Gln Glu Ser Arg 275 280 285 Thr Arg Lys Arg Arg Gly Ser Arg Val Ser Gln Asp Arg Asp Ser Glu 290 295 300 Gly His Ser Glu Asp Ser Glu Arg His Ser Gly Ser Ala Ser Arg Asn 305 310 315 320 His His Gly Ser Ala Trp Glu Gln Ser Arg Asp Gly Ser Arg His Pro 325 330 335 Arg Ser His Asp Glu Asp Arg Ala Ser His Gly His Ser Ala Asp Ser 340 345 350 Ser Arg Gln Ser Gly Thr Arg His Ala Glu Thr Ser Ser Arg Gly Gln 355 360 365 Thr Ala Ser Ser His Glu Gln Ala Arg Ser Ser Pro Gly Glu Arg His 370 375 380 Gly Ser Gly His Gln Gln Ser Ala Asp Ser Ser Arg His Ser Ala Thr 385 390 395 400 Gly Arg Gly Gln Ala Ser Ser Ala Val Ser Asp Arg Gly His Arg Gly 405 410 415 Ser Ser Gly Ser Gln Ala Ser Asp Ser Glu Gly His Ser Glu Asn Ser 420 425 430 Asp Thr Gln Ser Val Ser Gly His Gly Lys Ala Gly Leu Arg Gln Gln 435 440 445 Ser His Gln Glu Ser Thr Arg Gly Arg Ser Gly Glu Arg Ser Gly Arg 450 455 460 Ser Gly Ser Ser Leu Tyr Gln Val Ser Thr His Glu Gln Pro Asp Ser 465 470 475 480 Ala His Gly Arg Thr Gly Thr Ser Thr Gly Gly Arg Gln Gly Ser His 485 490 495 His Glu Gln Ala Arg Asp Ser Ser Arg His Ser Ala Ser Gln Glu Gly 500 505 510 Gln Asp Thr Ile Arg Gly His Pro Gly Ser Ser Arg Gly Gly Arg Gln 515 520 525 Gly Ser His His Glu Gln Ser Val Asn Arg Ser Gly His Ser Gly Ser 530 535 540 His His Ser His Thr Thr Ser Gln Gly Arg Ser Asp Ala Ser His Gly 545 550 555 560 Gln Ser Gly Ser Arg Ser Ala Ser Arg Gln Thr Arg Asn Glu Glu Gln 565 570 575 Ser Gly Asp Gly Thr Arg His Ser Gly Ser Arg His His Glu Ala Ser 580 585 590 Ser Gln Ala Asp Ser Ser Arg His Ser Gln Val Gly Gln Gly Gln Ser 595 600 605 Ser Gly Pro Arg Thr Ser Arg Asn Gln Gly Ser Ser Val Ser Gln Asp 610 615 620 Ser Asp Ser Gln Gly His Ser Glu Asp Ser Glu Arg Trp Ser Gly Ser 625 630 635 640 Ala Ser Arg Asn His His Gly Ser Ala Gln Glu Gln Ser Arg Asp Gly 645 650 655 Ser Arg His Pro Arg Ser His His Glu Asp Arg Ala Gly His Gly His 660 665 670 Ser Ala Asp Ser Ser Arg Lys Ser Gly Thr Arg His Thr Gln Asn Ser 675 680 685 Ser Ser Gly Gln Ala Ala Ser Ser His Glu Gln Ala Arg Ser Ser Ala 690 695 700 Gly Glu Arg His Gly Ser Arg His Gln Leu Gln Ser Ala Asp Ser Ser 705 710 715 720 Arg His Ser Gly Thr Gly His Gly Gln Ala Ser Ser Ala Val Arg Asp 725 730 735 Ser Gly His Arg Gly Ser Ser Gly Ser Gln Ala Thr Asp Ser Glu Gly 740 745 750 His Ser Glu Asp Ser Asp Thr Gln Ser Val Ser Gly His Gly Gln Ala 755 760 765 Gly His His Gln Gln Ser His Gln Glu Ser Ala Arg Asp Arg Ser Gly 770 775 780 Glu Arg Ser Arg Arg Ser Gly Ser Phe Leu Tyr Gln Val Ser Thr His 785 790 795 800 Lys Gln Ser Glu Ser Ser His Gly Trp Thr Gly Pro Ser Thr Gly Val 805 810 815 Arg Gln Gly Ser His His Glu Gln Ala Arg Asp Asn Ser Arg His Ser 820 825 830 Ala Ser Gln Asp Gly Gln Asp Thr Ile Arg Gly His Pro Gly Ser Ser 835 840 845 Arg Arg Gly Arg Gln Gly Ser His His Glu Gln Ser Val Asp Arg Ser 850 855 860 Gly His Ser Gly Ser His His Ser His Thr Thr Ser Gln Gly Arg Ser 865 870 875 880 Asp Ala Ser Arg Gly Gln Ser Gly Ser Arg Ser Ala Ser Arg Thr Thr 885 890 895 Arg Asn Glu Glu Gln Ser Arg Asp Gly Ser Arg His Ser Gly Ser Arg 900 905 910 His His Glu Ala Ser Ser His Ala Asp Ile Ser Arg His Ser Gln Ala 915 920 925 Gly Gln Gly Gln Ser Glu Gly Ser Arg Thr Ser Arg Arg Gln Gly Ser 930 935 940 Ser Val Ser Gln Asp Ser Asp Ser Glu Gly His Ser Glu Asp Ser Glu 945 950 955 960 Arg Trp Ser Gly Ser Ala Ser Arg Asn His Arg Gly Ser Ala Gln Glu 965 970 975 Gln Ser Arg His Gly Ser Arg His Pro Arg Ser His His Glu Asp Arg 980 985 990 Ala Gly His Gly His Ser Ala Asp Ser Ser Arg Gln Ser Gly Thr Pro 995 1000 1005 His Ala Glu Thr Ser Ser Gly Gly Gln Ala Ala Ser Ser His Glu 1010 1015 1020 Gln Ala Arg Ser Ser Pro Gly Glu Arg His Gly Ser Arg His Gln 1025 1030 1035 Gln Ser Ala Asp Ser Ser Arg His Ser Gly Ile Pro Arg Arg Gln 1040 1045 1050 Ala Ser Ser Ala Val Arg Asp Ser Gly His Trp Gly Ser Ser Gly 1055 1060 1065 Ser Gln Ala Ser Asp Ser Glu Gly His Ser Glu Glu Ser Asp Thr 1070 1075 1080 Gln Ser Val Ser Gly His Gly Gln Asp Gly Pro His Gln Gln Ser 1085 1090 1095 His Gln Glu Ser Ala Arg Asp Trp Ser Gly Gly Arg Ser Gly Arg 1100 1105 1110 Ser Gly Ser Phe Ile Tyr Gln Val Ser Thr His Glu Gln Ser Glu 1115 1120 1125 Ser Ala His Gly Arg Thr Arg Thr Ser Thr Gly Arg Arg Gln Gly 1130 1135 1140 Ser His His Glu Gln Ala Arg Asp Ser Ser Arg His Ser Ala Ser 1145 1150 1155 Gln Glu Gly Gln Asp Thr Ile Arg Ala His Pro Gly Ser Arg Arg 1160 1165 1170 Gly Gly Arg Gln Gly Ser His His Glu Gln Ser Val Asp Arg Ser 1175 1180 1185 Gly His Ser Gly Ser His His Ser His Thr Thr Ser Gln Gly Arg 1190 1195 1200 Ser Asp Ala Ser His Gly Gln Ser Gly Ser Arg Ser Ala Ser Arg 1205 1210 1215 Gln Thr Arg Lys Asp Lys Gln Ser Gly Asp Gly Ser Arg His Ser 1220 1225 1230 Gly Ser Arg His His Glu Ala Ala Ser Trp Ala Asp Ser Ser Arg 1235 1240 1245 His Ser Gln Val Gly Gln Glu Gln Ser Ser Gly Ser Arg Thr Ser 1250 1255 1260 Arg His Gln Gly Ser Ser Val Ser Gln Asp Ser Asp Ser Glu Arg 1265 1270 1275 His Ser Asp Asp Ser Glu Arg Leu Ser Gly Ser Ala Ser Arg Asn 1280 1285 1290 His His Gly Ser Ser Arg Glu Gln Ser Arg Asp Gly Ser Arg His 1295 1300 1305 Pro Gly Phe His Gln Glu Asp Arg Ala Ser His Gly His Ser Ala 1310 1315 1320 Asp Ser Ser Arg Gln Ser Gly Thr His His Thr Glu Ser Ser Ser 1325 1330 1335 His Gly Gln Ala Val Ser Ser His Glu Gln Ala Arg Ser Ser Pro 1340 1345 1350 Gly Glu Arg His Gly Ser Arg His Gln Gln Ser Ala Asp Ser Ser 1355 1360 1365 Arg His Ser Gly Ile Gly His Arg Gln Ala Ser Ser Ala Val Arg 1370 1375 1380 Asp Ser Gly His Arg Gly Ser Ser Gly Ser Gln Val Thr Asn Ser 1385 1390 1395 Glu Gly His Ser Glu Asp Ser Asp Thr Gln Ser Val Ser Ala His 1400 1405 1410 Gly Gln Ala Gly Pro His Gln Gln Ser His Lys Glu Ser Ala Arg 1415 1420 1425 Gly Gln Ser Gly Glu Ser Ser Gly Arg Ser Arg Ser Phe Leu Tyr 1430 1435 1440 Gln Val Ser Ser His Glu Gln Ser Glu Ser Thr His Gly Gln Thr 1445 1450 1455 Ala Pro Ser Thr Gly Gly Arg Gln Gly Ser Arg His Glu Gln Ala 1460 1465 1470 Arg Asn Ser Ser Arg His Ser Ala Ser Gln Asp Gly Gln Asp Thr 1475 1480 1485 Ile Arg Gly His Pro Gly Ser Ser Arg Gly Gly Arg Gln Gly Ser 1490 1495 1500 Tyr His Glu Gln Ser Val Asp Arg Ser Gly His Ser Gly Tyr His 1505 1510 1515 His Ser His Thr Thr Pro Gln Gly Arg Ser Asp Ala Ser His Gly 1520 1525 1530 Gln Ser Gly Pro Arg Ser Ala Ser Arg Gln Thr Arg Asn Glu Glu 1535 1540 1545 Gln Ser Gly Asp Gly Ser Arg His Ser Gly Ser Arg His His Glu 1550 1555 1560 Pro Ser Thr Arg Ala Gly Ser Ser Arg His Ser Gln Val Gly Gln 1565 1570 1575 Gly Glu Ser Ala Gly Ser Lys Thr Ser Arg Arg Gln Gly Ser Ser 1580 1585 1590 Val Ser Gln Asp Arg Asp Ser Glu Gly His Ser Glu Asp Ser Glu 1595 1600 1605 Arg Arg Ser Glu Ser Ala Ser Arg Asn His Tyr Gly Ser Ala Arg 1610 1615 1620 Glu Gln Ser Arg His Gly Ser Arg Asn Pro Arg Ser His Gln Glu 1625 1630 1635 Asp Arg Ala Ser His Gly His Ser Ala Glu Ser Ser Arg Gln Ser 1640 1645 1650 Gly Thr Arg His Ala Glu Thr Ser Ser Gly Gly Gln Ala Ala Ser 1655 1660 1665 Ser Gln Glu Gln Ala Arg Ser Ser Pro Gly Glu Arg His Gly Ser 1670 1675 1680 Arg His Gln Gln Ser Ala Asp Ser Ser Thr Asp Ser Gly Thr Gly 1685 1690 1695 Arg Arg Gln Asp Ser Ser Val Val Gly Asp Ser Gly Asn Arg Gly 1700 1705 1710 Ser Ser Gly Ser Gln Ala Ser Asp Ser Glu Gly His Ser Glu Glu 1715 1720 1725 Ser Asp Thr Gln Ser Val Ser Ala His Gly Gln Ala Gly Pro His 1730 1735 1740 Gln Gln Ser His Gln Glu Ser Thr Arg Gly Gln Ser Gly Glu Arg 1745 1750 1755 Ser Gly Arg Ser Gly Ser Phe Leu Tyr Gln Val Ser Thr His Glu 1760 1765 1770 Gln Ser Glu Ser Ala His Gly Arg Thr Gly Pro Ser Thr Gly Gly 1775 1780 1785 Arg Gln Arg Ser Arg His Glu Gln Ala Arg Asp Ser Ser Arg His 1790 1795 1800 Ser Ala Ser Gln Glu Gly Gln Asp Thr Ile Arg Gly His Pro Gly 1805 1810 1815 Ser Ser Arg Gly Gly Arg Gln Gly Ser His Tyr Glu Gln Ser Val 1820 1825 1830 Asp Ser Ser Gly His Ser Gly Ser His His Ser His Thr Thr Ser 1835 1840 1845 Gln Glu Arg Ser Asp Val Ser Arg Gly Gln Ser Gly Ser Arg Ser 1850 1855 1860 Val Ser Arg Gln Thr Arg Asn Glu Lys Gln Ser Gly Asp Gly Ser 1865 1870 1875 Arg His Ser Gly Ser Arg His His Glu Ala Ser Ser Arg Ala Asp 1880 1885 1890 Ser Ser Arg His Ser Gln Val Gly Gln Gly Gln Ser Ser Gly Pro 1895 1900 1905 Arg Thr Ser Arg Asn Gln Gly Ser Ser Val Ser Gln Asp Ser Asp 1910 1915 1920 Ser Gln Gly His Ser Glu Asp Ser Glu Arg Trp Ser Gly Ser Ala 1925 1930 1935 Ser Arg Asn His Leu Gly Ser Ala Trp Glu Gln Ser Arg Asp Gly 1940 1945 1950 Ser Arg His Pro Gly Ser His His Glu Asp Arg Ala Gly His Gly 1955 1960 1965 His Ser Ala Asp Ser Ser Arg Gln Ser Gly Thr Arg His Thr Glu 1970 1975 1980 Ser Ser Ser Arg Gly Gln Ala Ala Ser Ser His Glu Gln Ala Arg 1985 1990 1995 Ser Ser Ala Gly Glu Arg His Gly Ser His His Gln Leu Gln Ser 2000 2005 2010 Ala Asp Ser Ser Arg His Ser Gly Ile Gly His Gly Gln Ala Ser 2015 2020 2025 Ser Ala Val Arg Asp Ser Gly His Arg Gly Tyr Ser Gly Ser Gln 2030 2035 2040 Ala Ser Asp Ser Glu Gly His Ser Glu Asp Ser Asp Thr Gln Ser 2045 2050 2055 Val Ser Ala Gln Gly Lys Ala Gly Pro His Gln Gln Ser His Lys 2060 2065 2070 Glu Ser Ala Arg Gly Gln Ser Gly Glu Ser Ser Gly Arg Ser Gly 2075 2080 2085 Ser Phe Leu Tyr Gln Val Ser Thr His Glu Gln Ser Glu Ser Thr 2090 2095 2100 His Gly Gln Ser Ala Pro Ser Thr Gly Gly Arg Gln Gly Ser His 2105 2110 2115 Tyr Asp Gln Ala Gln Asp Ser Ser Arg His Ser Ala Ser Gln Glu 2120 2125 2130 Gly Gln Asp Thr Ile Arg Gly His Pro Gly Pro Ser Arg Gly Gly 2135 2140 2145 Arg Gln Gly Ser His Gln Glu Gln Ser Val Asp Arg Ser Gly His 2150 2155 2160 Ser Gly Ser His His Ser His Thr Thr Ser Gln Gly Arg Ser Asp 2165 2170 2175 Ala Ser Arg Gly Gln Ser Gly Ser Arg Ser Ala Ser Arg Lys Thr 2180 2185 2190 Tyr Asp Lys Glu Gln Ser Gly Asp Gly Ser Arg His Ser Gly Ser 2195 2200 2205 His His His Glu Ala Ser Ser Trp Ala Asp Ser Ser Arg His Ser 2210 2215 2220 Leu Val Gly Gln Gly Gln Ser Ser Gly Pro Arg Thr Ser Arg Pro 2225 2230 2235 Arg Gly Ser Ser Val Ser Gln Asp Ser Asp Ser Glu Gly His Ser 2240 2245 2250 Glu Asp Ser Glu Arg Arg Ser Gly Ser Ala Ser Arg Asn His His 2255 2260 2265 Gly Ser Ala Gln Glu Gln Ser Arg Asp Gly Ser Arg His Pro Arg 2270 2275 2280 Ser His His Glu Asp Arg Ala Gly His Gly His Ser Ala Glu Ser 2285 2290 2295 Ser Arg Gln Ser Gly Thr His His Ala Glu Asn Ser Ser Gly Gly 2300 2305 2310 Gln Ala Ala Ser Ser His Glu Gln Ala Arg Ser Ser Ala Gly Glu 2315 2320 2325 Arg His Gly Ser His His Gln Gln Ser Ala Asp Ser Ser Arg His 2330 2335 2340 Ser Gly Ile Gly His Gly Gln Ala Ser Ser Ala Val Arg Asp Ser 2345 2350 2355 Gly His Arg Gly Ser Ser Gly Ser Gln Ala Ser Asp Ser Glu Gly 2360 2365 2370 His Ser Glu Asp Ser Asp Thr Gln Ser Val Ser Ala His Gly Gln 2375 2380 2385 Ala Gly Pro His Gln Gln Ser His Gln Glu Ser Thr Arg Gly Arg 2390 2395 2400 Ser Ala Gly Arg Ser Gly Arg Ser Gly Ser Phe Leu Tyr Gln Val 2405 2410 2415 Ser Thr His Glu Gln Ser Glu Ser Ala His Gly Arg Thr Gly Thr 2420 2425 2430 Ser Thr Gly Gly Arg Gln Gly Ser His His Lys Gln Ala Arg Asp 2435 2440 2445 Ser Ser Arg His Ser Thr Ser Gln Glu Gly Gln Asp Thr Ile His 2450 2455 2460 Gly His Pro Gly Ser Ser Ser Gly Gly Arg Gln Gly Ser His Tyr 2465 2470 2475 Glu Gln Leu Val Asp Arg Ser Gly His Ser Gly Ser His His Ser 2480 2485 2490 His Thr Thr Ser Gln Gly Arg Ser Asp Ala Ser His Gly His Ser 2495 2500 2505 Gly Ser Arg Ser Ala Ser Arg Gln Thr Arg Asn Asp Glu Gln Ser 2510 2515 2520 Gly Asp Gly Ser Arg His Ser Gly Ser Arg His His Glu Ala Ser 2525 2530 2535 Ser Arg Ala Asp Ser Ser Gly His Ser Gln Val Gly Gln Gly Gln 2540 2545 2550 Ser Glu Gly Pro Arg Thr Ser Arg Asn Trp Gly Ser Ser Phe Ser 2555 2560 2565 Gln Asp Ser Asp Ser Gln Gly His Ser Glu Asp Ser Glu Arg Trp 2570 2575 2580 Ser Gly Ser Ala Ser Arg Asn His His Gly Ser Ala Gln Glu Gln 2585 2590 2595 Leu Arg Asp Gly Ser Arg His Pro Arg Ser His Gln Glu Asp Arg 2600 2605 2610 Ala Gly His Gly His Ser Ala Asp Ser Ser Arg Gln Ser Gly Thr 2615 2620 2625 Arg His Thr Gln Thr Ser Ser Gly Gly Gln Ala Ala Ser Ser His 2630 2635 2640 Glu Gln Ala Arg Ser Ser Ala Gly Glu Arg His Gly Ser His His 2645 2650 2655 Gln Gln Ser Ala Asp Ser Ser Arg His Ser Gly Ile Gly His Gly 2660 2665 2670 Gln Ala Ser Ser Ala Val Arg Asp Ser Gly His Arg Gly Tyr Ser 2675 2680 2685 Gly Ser Gln Ala Ser Asp Asn Glu Gly His Ser Glu Asp Ser Asp 2690 2695 2700 Thr Gln Ser Val Ser Ala His Gly Gln Ala Gly Ser His Gln Gln 2705 2710 2715 Ser His Gln Glu Ser Ala Arg Gly Arg Ser Gly Glu Thr Ser Gly 2720 2725 2730 His Ser Gly Ser Phe Leu Tyr Gln Val Ser Thr His Glu Gln Ser 2735 2740 2745 Glu Ser Ser His Gly Trp Thr Gly Pro Ser Thr Arg Gly Arg Gln 2750 2755 2760 Gly Ser Arg His Glu Gln Ala Gln Asp Ser Ser Arg His Ser Ala 2765 2770 2775 Ser Gln Asp Gly Gln Asp Thr Ile Arg Gly His Pro Gly Ser Ser 2780 2785 2790 Arg Gly Gly Arg Gln Gly Tyr His His Glu His Ser Val Asp Ser 2795 2800 2805 Ser Gly His Ser Gly Ser His His Ser His Thr Thr Ser Gln Gly 2810 2815 2820 Arg Ser Asp Ala Ser Arg Gly Gln Ser Gly Ser Arg Ser Ala Ser 2825 2830 2835 Arg Thr Thr Arg Asn Glu Glu Gln Ser Gly Asp Gly Ser Arg His 2840 2845 2850 Ser Gly Ser Arg His His Glu Ala Ser Thr His Ala Asp Ile Ser 2855 2860 2865 Arg His Ser Gln Ala Val Gln Gly Gln Ser Glu Gly Ser Arg Arg 2870 2875 2880 Ser Arg Arg Gln Gly Ser Ser Val Ser Gln Asp Ser Asp Ser Glu 2885 2890 2895 Gly His Ser Glu Asp Ser Glu Arg Trp Ser Gly Ser Ala Ser Arg 2900 2905 2910 Asn His His Gly Ser Ala Gln Glu Gln Leu Arg Asp Gly Ser Arg 2915 2920 2925 His Pro Arg Ser His Gln Glu Asp Arg Ala Gly His Gly His Ser 2930 2935 2940 Ala Asp Ser Ser Arg Gln Ser Gly Thr Arg His Thr Gln Thr Ser 2945 2950 2955 Ser Gly Gly Gln Ala Ala Ser Ser His Glu Gln Ala Arg Ser Ser 2960 2965 2970 Ala Gly Glu Arg His Gly Ser His His Gln Gln Ser Ala Asp Ser 2975 2980 2985 Ser Arg His Ser Gly Ile Gly His Gly Gln Ala Ser Ser Ala Val 2990 2995 3000 Arg Asp Ser Gly His Arg Gly Tyr Ser Gly Ser Gln Ala Ser Asp 3005 3010 3015 Asn Glu Gly His Ser Glu Asp Ser Asp Thr Gln Ser Val Ser Ala 3020 3025 3030 His Gly Gln Ala Gly Ser His Gln Gln Ser His Gln Glu Ser Ala 3035 3040 3045 Arg Gly Arg Ser Gly Glu Thr Ser Gly His Ser Gly Ser Phe Leu 3050 3055 3060 Tyr Gln Val Ser Thr His Glu Gln Ser Glu Ser Ser His Gly Trp 3065 3070 3075 Thr Gly Pro Ser Thr Arg Gly Arg Gln Gly Ser Arg His Glu Gln 3080 3085 3090 Ala Gln Asp Ser Ser Arg His Ser Ala Ser Gln Tyr Gly Gln Asp 3095 3100 3105 Thr Ile Arg Gly His Pro Gly Ser Ser Arg Gly Gly Arg Gln Gly 3110 3115 3120 Tyr His His Glu His Ser Val Asp Ser Ser Gly His Ser Gly Ser 3125 3130 3135 His His Ser His Thr Thr Ser Gln Gly Arg Ser Asp Ala Ser Arg 3140 3145 3150 Gly Gln Ser Gly Ser Arg Ser Ala Ser Arg Thr Thr Arg Asn Glu 3155 3160 3165 Glu Gln Ser Gly Asp Ser Ser Arg His Ser Val Ser Arg His His 3170 3175 3180 Glu Ala Ser Thr His Ala Asp Ile Ser Arg His Ser Gln Ala Val 3185 3190 3195 Gln Gly Gln Ser Glu Gly Ser Arg Arg Ser Arg Arg Gln Gly Ser 3200 3205 3210 Ser Val Ser Gln Asp Ser Asp Ser Glu Gly His Ser Glu Asp Ser 3215 3220 3225 Glu Arg Trp Ser Gly Ser Ala Ser Arg Asn His Arg Gly Ser Val 3230 3235 3240 Gln Glu Gln Ser Arg His Gly Ser Arg His Pro Arg Ser His His 3245 3250 3255 Glu Asp Arg Ala Gly His Gly His Ser Ala Asp Arg Ser Arg Gln 3260 3265 3270 Ser Gly Thr Arg His Ala Glu Thr Ser Ser Gly Gly Gln Ala Ala 3275 3280 3285 Ser Ser His Glu Gln Ala Arg Ser Ser Pro Gly Glu Arg His Gly 3290 3295 3300 Ser Arg His Gln Gln Ser Ala Asp Ser Ser Arg His Ser Gly Ile 3305 3310 3315 Pro Arg Gly Gln Ala Ser Ser Ala Val Arg Asp Ser Arg His Trp 3320 3325 3330 Gly Ser Ser Gly Ser Gln Ala Ser Asp Ser Glu Gly His Ser Glu 3335 3340 3345 Glu Ser Asp Thr Gln Ser Val Ser Gly His Gly Gln Ala Gly Pro 3350 3355 3360 His Gln Gln Ser His Gln Glu Ser Ala Arg Asp Arg Ser Gly Gly 3365 3370 3375 Arg Ser Gly Arg Ser Gly Ser Phe Leu Tyr Gln Val Ser Thr His 3380 3385 3390 Glu Gln Ser Glu Ser Ala His Gly Arg Thr Arg Thr Ser Thr Gly 3395 3400 3405 Arg Arg Gln Gly Ser His His Glu Gln Ala Arg Asp Ser Ser Arg 3410 3415 3420 His Ser Ala Ser Gln Glu Gly Gln Asp Thr Ile Arg Gly His Pro 3425 3430 3435 Gly Ser Ser Arg Arg Gly Arg Gln Gly Ser His Tyr Glu Gln Ser 3440 3445 3450 Val Asp Arg Ser Gly His Ser Gly Ser His His Ser His Thr Thr 3455 3460 3465 Ser Gln Gly Arg Ser Asp Ala Ser Arg Gly Gln Ser Gly Ser Arg 3470 3475 3480 Ser Ala Ser Arg Gln Thr Arg Asn Asp Glu Gln Ser Gly Asp Gly 3485 3490 3495 Ser Arg His Ser Trp Ser His His His Glu Ala Ser Thr Gln Ala 3500 3505 3510 Asp Ser Ser Arg His Ser Gln Ser Gly Gln Gly Gln Ser Ala Gly 3515 3520 3525 Pro Arg Thr Ser Arg Asn Gln Gly Ser Ser Val Ser Gln Asp Ser 3530 3535 3540 Asp Ser Gln Gly His Ser Glu Asp Ser Glu Arg Trp Ser Gly Ser 3545 3550 3555 Ala Ser Arg Asn His Arg Gly Ser Ala Gln Glu Gln Ser Arg Asp 3560 3565 3570 Gly Ser Arg His Pro Thr Ser His His Glu Asp Arg Ala Gly His 3575 3580 3585 Gly His Ser Ala Glu Ser Ser Arg Gln Ser Gly Thr His His Ala 3590 3595 3600 Glu Asn Ser Ser Gly Gly Gln Ala Ala Ser Ser His Glu Gln Ala 3605 3610 3615 Arg Ser Ser Ala Gly Glu Arg His Gly Ser His His Gln Gln Ser 3620 3625 3630 Ala Asp Ser Ser Arg His Ser Gly Ile Gly His Gly Gln Ala Ser 3635 3640 3645 Ser Ala Val Arg Asp Ser Gly His Arg Gly Ser Ser Gly Ser Gln 3650 3655 3660 Ala Ser Asp Ser Glu Gly His Ser Glu Asp Ser Asp Thr Gln Ser 3665 3670 3675 Val Ser Ala His Gly Gln Ala Gly Pro His Gln Gln Ser His Gln 3680 3685 3690 Glu Ser Thr Arg Gly Arg Ser Ala Gly Arg Ser Gly Arg Ser Gly 3695 3700 3705 Ser Phe Leu Tyr Gln Val Ser Thr His Glu Gln Ser Glu Ser Ala 3710 3715 3720 His Gly Arg Ala Gly Pro Ser Thr Gly Gly Arg Gln Gly Ser Arg 3725 3730 3735 His Glu Gln Ala Arg Asp Ser Ser Arg His Ser Ala Ser Gln Glu 3740 3745 3750 Gly Gln Asp Thr Ile Arg Gly His Pro Gly Ser Arg Arg Gly Gly 3755 3760 3765 Arg Gln Gly Ser Tyr His Glu Gln Ser Val Asp Arg Ser Gly His 3770 3775 3780 Ser Gly Ser His His Ser His Thr Thr Ser Gln Gly Arg Ser Asp 3785 3790 3795 Ala Ser His Gly Gln Ser Gly Ser Arg Ser Ala Ser Arg Glu Thr 3800 3805 3810 Arg Asn Glu Glu Gln Ser Gly Asp Gly Ser Arg His Ser Gly Ser 3815 3820 3825 Arg His His Glu Ala Ser Thr Gln Ala Asp Ser Ser Arg His Ser 3830 3835 3840 Gln Ser Gly Gln Gly Glu Ser Ala Gly Ser Arg Arg Ser Arg Arg 3845 3850 3855 Gln Gly Ser Ser Val Ser Gln Asp Ser Asp Ser Glu Ala Tyr Pro 3860 3865 3870 Glu Asp Ser Glu Arg Arg Ser Glu Ser Ala Ser Arg Asn His His 3875 3880 3885 Gly Ser Ser Arg Glu Gln Ser Arg Asp Gly Ser Arg His Pro Gly 3890 3895 3900 Ser Ser His Arg Asp Thr Ala Ser His Val Gln Ser Ser Pro Val 3905 3910 3915 Gln Ser Asp Ser Ser Thr Ala Lys Glu His Gly His Phe Ser Ser 3920 3925 3930 Leu Ser Gln Asp Ser Ala Tyr His Ser Gly Ile Gln Ser Arg Gly 3935 3940 3945 Ser Pro His Ser Ser Ser Ser Tyr His Tyr Gln Ser Glu Gly Thr 3950 3955 3960 Glu Arg Gln Lys Gly Gln Ser Gly Leu Val Trp Arg His Gly Ser 3965 3970 3975 Tyr Gly Ser Ala Asp Tyr Asp Tyr Gly Glu Ser Gly Phe Arg His 3980 3985 3990 Ser Gln His Gly Ser Val Ser Tyr Asn Ser Asn Pro Val Val Phe 3995 4000 4005 Lys Glu Arg Ser Asp Ile Cys Lys Ala Ser Ala Phe Gly Lys Asp 4010 4015 4020 His Pro Arg Tyr Tyr Ala Thr Tyr Ile Asn Lys Asp Pro Gly Leu 4025 4030 4035 Cys Gly His Ser Ser Asp Ile Ser Lys Gln Leu Gly Phe Ser Gln 4040 4045 4050 Ser Gln Arg Tyr Tyr Tyr Tyr Glu 4055 4060 <210> 2 <211> 346 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 2 Met Gln Ser Gly Glu Ser Ser Gly Arg Ser Arg Ser Phe Leu Tyr Gln 1 5 10 15 Val Ser Ser His Glu Gln Ser Glu Ser Thr His Gly Gln Thr Ala Pro 20 25 30 Ser Thr Gly Gly Arg Gln Gly Ser Arg His Glu Gln Ala Arg Asn Ser 35 40 45 Ser Arg His Ser Ala Ser Gln Asp Gly Gln Asp Thr Ile Arg Gly His 50 55 60 Pro Gly Ser Ser Arg Gly Gly Arg Gln Gly Ser Tyr His Glu Gln Ser 65 70 75 80 Val Asp Arg Ser Gly His Ser Gly Tyr His His Ser His Thr Thr Pro 85 90 95 Gln Gly Arg Ser Asp Ala Ser His Gly Gln Ser Gly Pro Arg Ser Ala 100 105 110 Ser Arg Gln Thr Arg Asn Glu Glu Gln Ser Gly Asp Gly Ser Arg His 115 120 125 Ser Gly Ser Arg His His Glu Pro Ser Thr Arg Ala Gly Ser Ser Arg 130 135 140 His Ser Gln Val Gly Gln Gly Glu Ser Ala Gly Ser Lys Thr Ser Arg 145 150 155 160 Arg Gln Gly Ser Ser Val Ser Gln Asp Arg Asp Ser Glu Gly His Ser 165 170 175 Glu Asp Ser Glu Arg Arg Ser Glu Ser Ala Ser Arg Asn His Tyr Gly 180 185 190 Ser Ala Arg Glu Gln Ser Arg His Gly Ser Arg Asn Pro Arg Ser His 195 200 205 Gln Glu Asp Arg Ala Ser His Gly His Ser Ala Glu Ser Ser Arg Gln 210 215 220 Ser Gly Thr Arg His Ala Glu Thr Ser Ser Gly Gly Gln Ala Ala Ser 225 230 235 240 Ser Gln Glu Gln Ala Arg Ser Ser Pro Gly Glu Arg His Gly Ser Arg 245 250 255 His Gln Gln Ser Ala Asp Ser Ser Thr Asp Ser Gly Thr Gly Arg Arg 260 265 270 Gln Asp Ser Ser Val Val Gly Asp Ser Gly Asn Arg Gly Ser Ser Gly 275 280 285 Ser Gln Ala Ser Asp Ser Glu Gly His Ser Glu Glu Ser Asp Thr Gln 290 295 300 Ser Val Ser Ala His Gly Gln Ala Gly Pro His Gln Gln Ser His Gln 305 310 315 320 Glu Ser Thr Arg Gly Gln Ser Gly Glu Arg Ser Gly Arg Ser Gly Ser 325 330 335 Phe Leu Tyr Gln Val Ser Thr His Glu Gln 340 345 <210> 3 <211> 356 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 3 Met Gln Ser Gly Glu Ser Ser Gly Arg Ser Arg Ser Phe Leu Tyr Gln 1 5 10 15 Val Ser Ser His Glu Gln Ser Glu Ser Thr His Gly Gln Thr Ala Pro 20 25 30 Ser Thr Gly Gly Arg Gln Gly Ser Arg His Glu Gln Ala Arg Asn Ser 35 40 45 Ser Arg His Ser Ala Ser Gln Asp Gly Gln Asp Thr Ile Arg Gly His 50 55 60 Pro Gly Ser Ser Arg Gly Gly Arg Gln Gly Ser Tyr His Glu Gln Ser 65 70 75 80 Val Asp Arg Ser Gly His Ser Gly Tyr His His Ser His Thr Thr Pro 85 90 95 Gln Gly Arg Ser Asp Ala Ser His Gly Gln Ser Gly Pro Arg Ser Ala 100 105 110 Ser Arg Gln Thr Arg Asn Glu Glu Gln Ser Gly Asp Gly Ser Arg His 115 120 125 Ser Gly Ser Arg His His Glu Pro Ser Thr Arg Ala Gly Ser Ser Arg 130 135 140 His Ser Gln Val Gly Gln Gly Glu Ser Ala Gly Ser Lys Thr Ser Arg 145 150 155 160 Arg Gln Gly Ser Ser Val Ser Gln Asp Arg Asp Ser Glu Gly His Ser 165 170 175 Glu Asp Ser Glu Arg Arg Ser Glu Ser Ala Ser Arg Asn His Tyr Gly 180 185 190 Ser Ala Arg Glu Gln Ser Arg His Gly Ser Arg Asn Pro Arg Ser His 195 200 205 Gln Glu Asp Arg Ala Ser His Gly His Ser Ala Glu Ser Ser Arg Gln 210 215 220 Ser Gly Thr Arg His Ala Glu Thr Ser Ser Gly Gly Gln Ala Ala Ser 225 230 235 240 Ser Gln Glu Gln Ala Arg Ser Ser Pro Gly Glu Arg His Gly Ser Arg 245 250 255 His Gln Gln Ser Ala Asp Ser Ser Thr Asp Ser Gly Thr Gly Arg Arg 260 265 270 Gln Asp Ser Ser Val Val Gly Asp Ser Gly Asn Arg Gly Ser Ser Gly 275 280 285 Ser Gln Ala Ser Asp Ser Glu Gly His Ser Glu Glu Ser Asp Thr Gln 290 295 300 Ser Val Ser Ala His Gly Gln Ala Gly Pro His Gln Gln Ser His Gln 305 310 315 320 Glu Ser Thr Arg Gly Gln Ser Gly Glu Arg Ser Gly Arg Ser Gly Ser 325 330 335 Phe Leu Tyr Gln Val Ser Thr His Glu Gln Arg Met Arg Arg Met Arg 340 345 350 Arg Met Arg Arg 355 <210> 4 <211> 337 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 4 Met Glu Ser Thr His Gly Gln Thr Ala Pro Ser Thr Gly Gly Arg Gln 1 5 10 15 Gly Ser Arg His Glu Gln Ala Arg Asn Ser Ser Arg His Ser Ala Ser 20 25 30 Gln Asp Gly Gln Asp Thr Ile Arg Gly His Pro Gly Ser Ser Arg Gly 35 40 45 Gly Arg Gln Gly Ser Tyr His Glu Gln Ser Val Asp Arg Ser Gly His 50 55 60 Ser Gly Tyr His His Ser His Thr Thr Pro Gln Gly Arg Ser Asp Ala 65 70 75 80 Ser His Gly Gln Ser Gly Pro Arg Ser Ala Ser Arg Gln Thr Arg Asn 85 90 95 Glu Glu Gln Ser Gly Asp Gly Ser Arg His Ser Gly Ser Arg His His 100 105 110 Glu Pro Ser Thr Arg Ala Gly Ser Ser Arg His Ser Gln Val Gly Gln 115 120 125 Gly Glu Ser Ala Gly Ser Lys Thr Ser Arg Arg Gln Gly Ser Ser Val 130 135 140 Ser Gln Asp Arg Asp Ser Glu Gly His Ser Glu Asp Ser Glu Arg Arg 145 150 155 160 Ser Glu Ser Ala Ser Arg Asn His Tyr Gly Ser Ala Arg Glu Gln Ser 165 170 175 Arg His Gly Ser Arg Asn Pro Arg Ser His Gln Glu Asp Arg Ala Ser 180 185 190 His Gly His Ser Ala Glu Ser Ser Arg Gln Ser Gly Thr Arg His Ala 195 200 205 Glu Thr Ser Ser Gly Gly Gln Ala Ala Ser Ser Gln Glu Gln Ala Arg 210 215 220 Ser Ser Pro Gly Glu Arg His Gly Ser Arg His Gln Gln Ser Ala Asp 225 230 235 240 Ser Ser Thr Asp Ser Gly Thr Gly Arg Arg Gln Asp Ser Ser Val Val 245 250 255 Gly Asp Ser Gly Asn Arg Gly Ser Ser Gly Ser Gln Ala Ser Asp Ser 260 265 270 Glu Gly His Ser Glu Glu Ser Asp Thr Gln Ser Val Ser Ala His Gly 275 280 285 Gln Ala Gly Pro His Gln Gln Ser His Gln Glu Ser Thr Arg Gly Gln 290 295 300 Ser Gly Glu Arg Ser Gly Arg Ser Gly Ser Phe Leu Tyr Gln Val Ser 305 310 315 320 Thr His Glu Gln Ser Glu Ser Arg Met Arg Arg Met Arg Arg Met Arg 325 330 335 Arg <210> 5 <211> 336 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 5 Met Arg Asn Glu Glu Gln Ser Gly Asp Gly Ser Arg His Ser Gly Ser 1 5 10 15 Arg His His Glu Pro Ser Thr Arg Ala Gly Ser Ser Arg His Ser Gln 20 25 30 Val Gly Gln Gly Glu Ser Ala Gly Ser Lys Thr Ser Arg Arg Gln Gly 35 40 45 Ser Ser Val Ser Gln Asp Arg Asp Ser Glu Gly His Ser Glu Asp Ser 50 55 60 Glu Arg Arg Ser Glu Ser Ala Ser Arg Asn His Tyr Gly Ser Ala Arg 65 70 75 80 Glu Gln Ser Arg His Gly Ser Arg Asn Pro Arg Ser His Gln Glu Asp 85 90 95 Arg Ala Ser His Gly His Ser Ala Glu Ser Ser Arg Gln Ser Gly Thr 100 105 110 Arg His Ala Glu Thr Ser Ser Gly Gly Gln Ala Ala Ser Ser Gln Glu 115 120 125 Gln Ala Arg Ser Ser Pro Gly Glu Arg His Gly Ser Arg His Gln Gln 130 135 140 Ser Ala Asp Ser Ser Thr Asp Ser Gly Thr Gly Arg Arg Gln Asp Ser 145 150 155 160 Ser Val Val Gly Asp Ser Gly Asn Arg Gly Ser Ser Gly Ser Gln Ala 165 170 175 Ser Asp Ser Glu Gly His Ser Glu Glu Ser Asp Thr Gln Ser Val Ser 180 185 190 Ala His Gly Gln Ala Gly Pro His Gln Gln Ser His Gln Glu Ser Thr 195 200 205 Arg Gly Gln Ser Gly Glu Arg Ser Gly Arg Ser Gly Ser Phe Leu Tyr 210 215 220 Gln Val Ser Thr His Glu Gln Ser Glu Ser Ala His Gly Arg Thr Gly 225 230 235 240 Pro Ser Thr Gly Gly Arg Gln Arg Ser Arg His Glu Gln Ala Arg Asp 245 250 255 Ser Ser Arg His Ser Ala Ser Gln Glu Gly Gln Asp Thr Ile Arg Gly 260 265 270 His Pro Gly Ser Ser Arg Gly Gly Arg Gln Gly Ser His Tyr Glu Gln 275 280 285 Ser Val Asp Ser Ser Gly His Ser Gly Ser His His Ser His Thr Thr 290 295 300 Ser Gln Glu Arg Ser Asp Val Ser Arg Gly Gln Ser Gly Ser Arg Ser 305 310 315 320 Val Ser Arg Gln Thr Arg Arg Met Arg Arg Met Arg Arg Met Arg Arg 325 330 335 <210> 6 <211> 244 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 6 Met Arg Asn Glu Glu Gln Ser Gly Asp Gly Ser Arg His Ser Gly Ser 1 5 10 15 Arg His His Glu Pro Ser Thr Arg Ala Gly Ser Ser Arg His Ser Gln 20 25 30 Val Gly Gln Gly Glu Ser Ala Gly Ser Lys Thr Ser Arg Arg Gln Gly 35 40 45 Ser Ser Val Ser Gln Asp Arg Asp Ser Glu Gly His Ser Glu Asp Ser 50 55 60 Glu Arg Arg Ser Glu Ser Ala Ser Arg Asn His Tyr Gly Ser Ala Arg 65 70 75 80 Glu Gln Ser Arg His Gly Ser Arg Asn Pro Arg Ser His Gln Glu Asp 85 90 95 Arg Ala Ser His Gly His Ser Ala Glu Ser Ser Arg Gln Ser Gly Thr 100 105 110 Arg His Ala Glu Thr Ser Ser Gly Gly Gln Ala Ala Ser Ser Gln Glu 115 120 125 Gln Ala Arg Ser Ser Pro Gly Glu Arg His Gly Ser Arg His Gln Gln 130 135 140 Ser Ala Asp Ser Ser Thr Asp Ser Gly Thr Gly Arg Arg Gln Asp Ser 145 150 155 160 Ser Val Val Gly Asp Ser Gly Asn Arg Gly Ser Ser Gly Ser Gln Ala 165 170 175 Ser Asp Ser Glu Gly His Ser Glu Glu Ser Asp Thr Gln Ser Val Ser 180 185 190 Ala His Gly Gln Ala Gly Pro His Gln Gln Ser His Gln Glu Ser Thr 195 200 205 Arg Gly Gln Ser Gly Glu Arg Ser Gly Arg Ser Gly Ser Phe Leu Tyr 210 215 220 Gln Val Ser Thr His Glu Gln Ser Glu Ser Arg Met Arg Arg Met Arg 225 230 235 240 Arg Met Arg Arg <210> 7 <400> 7 000 <210> 8 <211> 352 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 8 Met Arg Ser Arg Ser Phe Leu Tyr Gln Val Ser Ser His Glu Gln Ser 1 5 10 15 Glu Ser Thr His Gly Gln Thr Ala Pro Ser Thr Gly Gly Arg Gln Gly 20 25 30 Ser Arg His Glu Gln Ala Arg Asn Ser Ser Arg His Ser Ala Ser Gln 35 40 45 Asp Gly Gln Asp Thr Ile Arg Gly His Pro Gly Ser Ser Arg Gly Gly 50 55 60 Arg Gln Gly Ser Tyr His Glu Gln Ser Val Asp Arg Ser Gly His Ser 65 70 75 80 Gly Tyr His His Ser His Thr Thr Pro Gln Gly Arg Ser Asp Ala Ser 85 90 95 His Gly Gln Ser Gly Pro Arg Ser Ala Ser Arg Gln Thr Arg Asn Glu 100 105 110 Glu Gln Ser Gly Asp Gly Ser Arg His Ser Gly Ser Arg His His Glu 115 120 125 Ala Ser Thr Arg Ala Asp Ser Ser Arg His Ser Gln Val Gly Gln Gly 130 135 140 Gln Ser Ser Gly Ser Arg Thr Ser Arg Arg Gln Gly Ser Ser Val Ser 145 150 155 160 Gln Asp Ser Asp Ser Glu Gly His Ser Glu Asp Ser Glu Arg Arg Ser 165 170 175 Gly Ser Ala Ser Arg Asn His Tyr Gly Ser Ala Gln Glu Gln Ser Arg 180 185 190 Asp Gly Ser Arg His Pro Arg Ser His Gln Glu Asp Arg Ala Ser His 195 200 205 Gly His Ser Ala Glu Ser Ser Arg Gln Ser Gly Thr Arg His Ala Glu 210 215 220 Thr Ser Ser Gly Gly Gln Ala Ala Ser Ser His Glu Gln Ala Arg Ser 225 230 235 240 Ser Pro Gly Glu Arg His Gly Ser Arg His Gln Gln Ser Ala Asp Ser 245 250 255 Ser Arg His Ser Gly Ile Gly His Gly Gln Ala Ser Ser Ala Val Arg 260 265 270 Asp Ser Gly His Arg Gly Ser Ser Gly Ser Gln Ala Ser Asp Ser Glu 275 280 285 Gly His Ser Glu Asp Ser Asp Thr Gln Ser Val Ser Ala His Gly Gln 290 295 300 Ala Gly Pro His Gln Gln Ser His Gln Glu Ser Ala Arg Gly Arg Ser 305 310 315 320 Gly Glu Arg Ser Gly Arg Ser Gly Ser Phe Leu Tyr Gln Val Ser Thr 325 330 335 His Glu Gln Ser Glu Ser Arg Met Arg Arg Met Arg Arg Met Arg Arg 340 345 350 <210> 9 <211> 328 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <220> <221> MOD_RES <222> (1)..(1) <223> Any amino acid <220> <221> MOD_RES <222> (9)..(9) <223> Any amino acid <220> <221> MOD_RES <222> (11)..(11) <223> Any amino acid <220> <221> MOD_RES <222> (14)..(14) <223> Any amino acid <220> <221> MOD_RES <222> (17)..(17) <223> Any amino acid <220> <221> MOD_RES <222> (19)..(20) <223> Any amino acid <220> <221> MOD_RES <222> (23)..(24) <223> Any amino acid <220> <221> MOD_RES <222> (27)..(27) <223> Any amino acid <220> <221> MOD_RES <222> (30)..(31) <223> Any amino acid <220> <221> MOD_RES <222> (34)..(36) <223> Any amino acid <220> <221> MOD_RES <222> (41)..(41) <223> Any amino acid <220> <221> MOD_RES <222> (44)..(44) <223> Any amino acid <220> <221> MOD_RES <222> (55)..(58) <223> Any amino acid <220> <221> MOD_RES <222> (63)..(65) <223> Any amino acid <220> <221> MOD_RES <222> (67)..(68) <223> Any amino acid <220> <221> MOD_RES <222> (70)..(71) <223> Any amino acid <220> <221> MOD_RES <222> (77)..(77) <223> Any amino acid <220> <221> MOD_RES <222> (84)..(84) <223> Any amino acid <220> <221> MOD_RES <222> (86)..(86) <223> Any amino acid <220> <221> MOD_RES <222> (94)..(94) <223> Any amino acid <220> <221> MOD_RES <222> (97)..(97) <223> Any amino acid <220> <221> MOD_RES <222> (103)..(103) <223> Any amino acid <220> <221> MOD_RES <222> (105)..(108) <223> Any amino acid <220> <221> MOD_RES <222> (111)..(111) <223> Any amino acid <220> <221> MOD_RES <222> (113)..(113) <223> Any amino acid <220> <221> MOD_RES <222> (118)..(118) <223> Any amino acid <220> <221> MOD_RES <222> (124)..(125) <223> Any amino acid <220> <221> MOD_RES <222> (127)..(127) <223> Any amino acid <220> <221> MOD_RES <222> (129)..(130) <223> Any amino acid <220> <221> MOD_RES <222> (132)..(132) <223> Any amino acid <220> <221> MOD_RES <222> (135)..(136) <223> Any amino acid <220> <221> MOD_RES <222> (139)..(140) <223> Any amino acid <220> <221> MOD_RES <222> (142)..(142) <223> Any amino acid <220> <221> MOD_RES <222> (144)..(144) <223> Any amino acid <220> <221> MOD_RES <222> (146)..(146) <223> Any amino acid <220> <221> MOD_RES <222> (149)..(150) <223> Any amino acid <220> <221> MOD_RES <222> (154)..(154) <223> Any amino acid <220> <221> MOD_RES <222> (158)..(158) <223> Any amino acid <220> <221> MOD_RES <222> (161)..(162) <223> Any amino acid <220> <221> MOD_RES <222> (170)..(170) <223> Any amino acid <220> <221> MOD_RES <222> (172)..(172) <223> Any amino acid <220> <221> MOD_RES <222> (179)..(179) <223> Any amino acid <220> <221> MOD_RES <222> (182)..(183) <223> Any amino acid <220> <221> MOD_RES <222> (186)..(186) <223> Any amino acid <220> <221> MOD_RES <222> (188)..(188) <223> Any amino acid <220> <221> MOD_RES <222> (192)..(192) <223> Any amino acid <220> <221> MOD_RES <222> (194)..(195) <223> Any amino acid <220> <221> MOD_RES <222> (197)..(197) <223> Any amino acid <220> <221> MOD_RES <222> (202)..(202) <223> Any amino acid <220> <221> MOD_RES <222> (209)..(209) <223> Any amino acid <220> <221> MOD_RES <222> (216)..(216) <223> Any amino acid <220> <221> MOD_RES <222> (218)..(220) <223> Any amino acid <220> <221> MOD_RES <222> (223)..(223) <223> Any amino acid <220> <221> MOD_RES <222> (230)..(230) <223> Any amino acid <220> <221> MOD_RES <222> (237)..(237) <223> Any amino acid <220> <221> MOD_RES <222> (247)..(247) <223> Any amino acid <220> <221> MOD_RES <222> (254)..(255) <223> Any amino acid <220> <221> MOD_RES <222> (258)..(259) <223> Any amino acid <220> <221> MOD_RES <222> (261)..(261) <223> Any amino acid <220> <221> MOD_RES <222> (263)..(263) <223> Any amino acid <220> <221> MOD_RES <222> (268)..(268) <223> Any amino acid <220> <221> MOD_RES <222> (271)..(273) <223> Any amino acid <220> <221> MOD_RES <222> (275)..(275) <223> Any amino acid <220> <221> MOD_RES <222> (282)..(283) <223> Any amino acid <220> <221> MOD_RES <222> (298)..(298) <223> Any amino acid <220> <221> MOD_RES <222> (300)..(301) <223> Any amino acid <220> <221> MOD_RES <222> (303)..(303) <223> Any amino acid <220> <221> MOD_RES <222> (309)..(309) <223> Any amino acid <220> <221> MOD_RES <222> (312)..(312) <223> Any amino acid <220> <221> MOD_RES <222> (314)..(315) <223> Any amino acid <220> <221> MOD_RES <222> (318)..(319) <223> Any amino acid <220> <221> MOD_RES <222> (321)..(321) <223> Any amino acid <220> <221> MOD_RES <222> (324)..(324) <223> Any amino acid <400> 9 Xaa Leu Tyr Gln Val Ser Thr His Xaa Gln Xaa Asp Ser Xaa His Gly 1 5 10 15 Xaa Thr Xaa Xaa Ser Thr Xaa Xaa Arg Gln Xaa Ser His Xaa Xaa Gln 20 25 30 Ala Xaa Xaa Xaa Ser Arg His Ser Xaa Ser Gln Xaa Gly Gln Asp Thr 35 40 45 Ile Arg Gly His Pro Gly Xaa Xaa Xaa Xaa Gly Arg Gln Gly Xaa Xaa 50 55 60 Xa Glu Xa Xa Val Xa Xa Ser Gly His Ser Gly Xa His His Ser 65 70 75 80 His Thr Thr Xaa Gln Xaa Arg Ser Asp Ala Ser His Gly Xaa Ser Gly 85 90 95 No Arg Ser Ala Ser Arg No Thr No No No no no no no no no no no no no no no no no no no no no no no Gln no no no Asp 100 105 110 No Thr Arg His Ser No Ser Arg His Glu No Ser No Ala 115 120 125 No Ser No His Ser No Gly Gln No Ser No Gly 130 135 140 Arg No Ser Arg No Gly Ser Ser Ser Gln Asp No Asp Ser 145 150 155 160 Yes Yes His Ser Glu Asp Ser Glu Arg Yes Yes Yes Ala Ser Arg 165 170 175 Asn His No Gly Ser No Glu Gln No Arg No Gly Ser Arg No. 180 185 190 Pro Rock Rock His Rock Glu Asp Arg Ala Rock His Gly His Ser Ala Asp 195 200 205 No Ser Arg Lys Ser Gly Thr No His No Ser Ser No Gly 210 215 220 Gln Only Ser Not Glu Gln Only Arg Ser Not Gly Glu Arg 225 230 235 240 His Gly Ser Arg His Gln Xaa Gln Ser Ala Asp Ser Ser Xa Xaa Ser 245 250 255 Gly No. His No. Gln No. Ser Ser Ala Val No. Asp Ser No. 260 265 270 No Gly No Ser Gly Ser Gln Only Thr No No Glu Gly His Ser Glu 275 280 285 Asp Ser Asp Thr Gln Ser Val Ser Gly Xaa Gly Xaa Gly Xaa His 290,295,300 Gln Gln Ser His No Glu Ser No Arg No Ser Gly No Ser 305 310 315 320 When Arg Will Be When Phe Leu Tyr 325 <210> 10 <211> 416 <212> PRT <213> Homo sapiens <400> 10 Ala Gly Pro His Gln Gln Ser His Gln Glu Ser Thr Arg Gly Arg Ser 1 5 10 15 Ala Gly Arg Ser Gly Arg Ser Gly Ser Phe Leu Tyr Gln Val Ser Thr 20 25 30 His Glu Gln Ser Glu Ser Ala His Gly Arg Thr Gly Thr Ser Thr Gly 35 40 45 Gly Arg Gln Gly Ser His His Gln Gln Ala Arg Asp Ser Ser Arg His 50 55 60 Ser Thr Ser Gln Glu Gly Gln Asp Thr Ile His Gly His Arg Gly Ser 65 70 75 80 Ser Ser Gly Gly Arg Gln Gly Ser His Tyr Glu Gln Leu Val Asp Arg 85 90 95 Ser Gly His Ser Gly Ser His His Ser His Thr Thr Ser Gln Gly Arg 100 105 110 Ser Asp Ala Ser His Gly His Ser Gly Ser Arg Ser Ala Ser Arg Gln 115 120 125 Thr Arg Asn Asp Glu Gln Ser Gly Asp Gly Ser Arg His Ser Gly Ser 130 135 140 Arg His His Glu Ala Ser Ser Arg Ala Asp Ser Ser Gly His Ser Gln 145 150 155 160 Val Gly Gln Gly Gln Ser Glu Gly Pro Arg Thr Ser Arg Asn Trp Gly 165 170 175 Ser Ser Phe Ser Gln Asp Ser Asp Ser Gln Gly His Ser Glu Asp Ser 180 185 190 Glu Arg Trp Ser Gly Ser Ala Ser Arg Asn His His Gly Ser Ala Gln 195 200 205 Glu Gln Leu Arg Asp Gly Ser Arg His Pro Arg Ser His Gln Glu Asp 210 215 220 Arg Ala Gly His Gly His Ser Ala Asp Ser Ser Arg Gln Ser Gly Thr 225 230 235 240 Arg His Thr Gln Thr Ser Ser Gly Gly Gln Ala Ala Ser Ser His Glu 245 250 255 Gln Ala Arg Ser Ser Ala Gly Asp Arg His Gly Ser His His Gln Gln 260 265 270 Ser Ala Asp Ser Ser Arg His Ser Gly Ile Gly His Gly Gln Ala Ser 275 280 285 Ser Ala Val Arg Asp Ser Gly His Arg Gly Tyr Ser Gly Ser Gln Ala 290 295 300 Ser Asp Asn Glu Gly His Ser Glu Asp Ser Asp Thr Gln Ser Val Ser 305 310 315 320 Ala His Gly Gln Ala Gly Ser His Gln Gln Ser His Gln Glu Ser Ala 325 330 335 Arg Gly Arg Ser Gly Glu Thr Ser Gly His Ser Gly Ser Phe Leu Tyr 340 345 350 Gln Val Ser Thr His Glu Gln Ser Glu Ser Ser His Gly Trp Thr Gly 355 360 365 Pro Ser Thr Arg Gly Arg Gln Gly Ser Arg His Glu Gln Ala Gln Asp 370 375 380 Ser Ser Arg His Ser Ala Ser Gln Asp Gly Gln Asp Thr Ile Arg Gly 385 390 395 400 His Pro Gly Ser Ser Arg Gly Gly Arg Gln Gly Tyr His His Glu Gln 405 410 415 <210> 11 <211> 4061 <212> PRT <213> Homo sapiens <400> 11 Met Ser Thr Leu Leu Glu Asn Ile Phe Ala Ile Ile Asn Leu Phe Lys 1 5 10 15 Gln Tyr Ser Lys Lys Asp Lys Asn Thr Asp Thr Leu Ser Lys Lys Glu 20 25 30 Leu Lys Glu Leu Leu Glu Lys Glu Phe Arg Gln Ile Leu Lys Asn Pro 35 40 45 Asp Asp Pro Asp Met Val Asp Val Phe Met Asp His Leu Asp Ile Asp 50 55 60 His Asn Lys Lys Ile Asp Phe Thr Glu Phe Leu Leu Met Val Phe Lys 65 70 75 80 Leu Ala Gln Ala Tyr Tyr Glu Ser Thr Arg Lys Glu Asn Leu Pro Ile 85 90 95 Ser Gly His Lys His Arg Lys His Ser His His Asp Lys His Glu Asp 100 105 110 Asn Lys Gln Glu Glu Asn Lys Glu Asn Arg Lys Arg Pro Ser Ser Leu 115 120 125 Glu Arg Arg Asn Asn Arg Lys Gly Asn Lys Gly Arg Ser Lys Ser Pro 130 135 140 Arg Glu Thr Gly Gly Lys Arg His Glu Ser Ser Ser Glu Lys Lys Glu 145 150 155 160 Arg Lys Gly Tyr Ser Pro Thr His Arg Glu Glu Glu Tyr Gly Lys Asn 165 170 175 His His Asn Ser Ser Lys Lys Glu Lys Asn Lys Thr Glu Asn Thr Arg 180 185 190 Leu Gly Asp Asn Arg Lys Arg Leu Ser Glu Arg Leu Glu Glu Lys Glu 195 200 205 Asp Asn Glu Glu Gly Val Tyr Asp Tyr Glu Asn Thr Gly Arg Met Thr 210 215 220 Gln Lys Trp Ile Gln Ser Gly His Ile Ala Thr Tyr Tyr Thr Ile Gln 225 230 235 240 Asp Glu Ala Tyr Asp Thr Thr Asp Ser Leu Leu Glu Glu Asn Lys Ile 245 250 255 Tyr Glu Arg Ser Arg Ser Ser Asp Gly Lys Ser Ser Ser Gln Val Asn 260 265 270 Arg Ser Arg His Glu Asn Thr Ser Gln Val Pro Leu Gln Glu Ser Arg 275 280 285 Thr Arg Lys Arg Arg Gly Ser Arg Val Ser Gln Asp Arg Asp Ser Glu 290 295 300 Gly His Ser Glu Asp Ser Glu Arg His Ser Gly Ser Ala Ser Arg Asn 305 310 315 320 His His Gly Ser Ala Trp Glu Gln Ser Arg Asp Gly Ser Arg His Pro 325 330 335 Arg Ser His Asp Glu Asp Arg Ala Ser His Gly His Ser Ala Asp Ser 340 345 350 Ser Arg Gln Ser Gly Thr Arg His Ala Glu Thr Ser Ser Arg Gly Gln 355 360 365 Thr Ala Ser Ser His Glu Gln Ala Arg Ser Ser Pro Gly Glu Arg His 370 375 380 Gly Ser Gly His Gln Gln Ser Ala Asp Ser Ser Arg His Ser Ala Thr 385 390 395 400 Gly Arg Gly Gln Ala Ser Ser Ala Val Ser Asp Arg Gly His Arg Gly 405 410 415 Ser Ser Gly Ser Gln Ala Ser Asp Ser Glu Gly His Ser Glu Asn Ser 420 425 430 Asp Thr Gln Ser Val Ser Gly His Gly Lys Ala Gly Leu Arg Gln Gln 435 440 445 Ser His Gln Glu Ser Thr Arg Gly Arg Ser Gly Glu Arg Ser Gly Arg 450 455 460 Ser Gly Ser Ser Leu Tyr Gln Val Ser Thr His Glu Gln Pro Asp Ser 465 470 475 480 Ala His Gly Arg Thr Gly Thr Ser Thr Gly Gly Arg Gln Gly Ser His 485 490 495 His Glu Gln Ala Arg Asp Ser Ser Arg His Ser Ala Ser Gln Glu Gly 500 505 510 Gln Asp Thr Ile Arg Gly His Pro Gly Ser Ser Arg Gly Gly Arg Gln 515 520 525 Gly Ser His His Glu Gln Ser Val Asn Arg Ser Gly His Ser Gly Ser 530 535 540 His His Ser His Thr Thr Ser Gln Gly Arg Ser Asp Ala Ser His Gly 545 550 555 560 Gln Ser Gly Ser Arg Ser Ala Ser Arg Gln Thr Arg Asn Glu Glu Gln 565 570 575 Ser Gly Asp Gly Thr Arg His Ser Gly Ser Arg His His Glu Ala Ser 580 585 590 Ser Gln Ala Asp Ser Ser Arg His Ser Gln Val Gly Gln Gly Gln Ser 595 600 605 Ser Gly Pro Arg Thr Ser Arg Asn Gln Gly Ser Ser Val Ser Gln Asp 610 615 620 Ser Asp Ser Gln Gly His Ser Glu Asp Ser Glu Arg Trp Ser Gly Ser 625 630 635 640 Ala Ser Arg Asn His His Gly Ser Ala Gln Glu Gln Ser Arg Asp Gly 645 650 655 Ser Arg His Pro Arg Ser His His Glu Asp Arg Ala Gly His Gly His 660 665 670 Ser Ala Asp Ser Ser Arg Lys Ser Gly Thr Arg His Thr Gln Asn Ser 675 680 685 Ser Ser Gly Gln Ala Ala Ser Ser His Glu Gln Ala Arg Ser Ser Ala 690 695 700 Gly Glu Arg His Gly Ser Arg His Gln Leu Gln Ser Ala Asp Ser Ser 705 710 715 720 Arg His Ser Gly Thr Gly His Gly Gln Ala Ser Ser Ala Val Arg Asp 725 730 735 Ser Gly His Arg Gly Ser Ser Gly Ser Gln Ala Thr Asp Ser Glu Gly 740 745 750 His Ser Glu Asp Ser Asp Thr Gln Ser Val Ser Gly His Gly Gln Ala 755 760 765 Gly His His Gln Gln Ser His Gln Glu Ser Ala Arg Asp Arg Ser Gly 770 775 780 Glu Arg Ser Arg Arg Ser Gly Ser Phe Leu Tyr Gln Val Ser Thr His 785 790 795 800 Lys Gln Ser Glu Ser Ser His Gly Trp Thr Gly Pro Ser Thr Gly Val 805 810 815 Arg Gln Gly Ser His His Glu Gln Ala Arg Asp Asn Ser Arg His Ser 820 825 830 Ala Ser Gln Asp Gly Gln Asp Thr Ile Arg Gly His Pro Gly Ser Ser 835 840 845 Arg Arg Gly Arg Gln Gly Ser His His Glu Gln Ser Val Asp Arg Ser 850 855 860 Gly His Ser Gly Ser His His Ser His Thr Thr Ser Gln Gly Arg Ser 865 870 875 880 Asp Ala Ser Arg Gly Gln Ser Gly Ser Arg Ser Ala Ser Arg Thr Thr 885 890 895 Arg Asn Glu Glu Gln Ser Arg Asp Gly Ser Arg His Ser Gly Ser Arg 900 905 910 His His Glu Ala Ser Ser His Ala Asp Ile Ser Arg His Ser Gln Ala 915 920 925 Gly Gln Gly Gln Ser Glu Gly Ser Arg Thr Ser Arg Arg Gln Gly Ser 930 935 940 Ser Val Ser Gln Asp Ser Asp Ser Glu Gly His Ser Glu Asp Ser Glu 945 950 955 960 Arg Trp Ser Gly Ser Ala Ser Arg Asn His Arg Gly Ser Ala Gln Glu 965 970 975 Gln Ser Arg His Gly Ser Arg His Pro Arg Ser His His Glu Asp Arg 980 985 990 Ala Gly His Gly His Ser Ala Asp Ser Ser Arg Gln Ser Gly Thr Pro 995 1000 1005 His Ala Glu Thr Ser Ser Gly Gly Gln Ala Ala Ser Ser His Glu 1010 1015 1020 Gln Ala Arg Ser Ser Pro Gly Glu Arg His Gly Ser Arg His Gln 1025 1030 1035 Gln Ser Ala Asp Ser Ser Arg His Ser Gly Ile Pro Arg Arg Gln 1040 1045 1050 Ala Ser Ser Ala Val Arg Asp Ser Gly His Trp Gly Ser Ser Gly 1055 1060 1065 Ser Gln Ala Ser Asp Ser Glu Gly His Ser Glu Glu Ser Asp Thr 1070 1075 1080 Gln Ser Val Ser Gly His Gly Gln Asp Gly Pro His Gln Gln Ser 1085 1090 1095 His Gln Glu Ser Ala Arg Asp Trp Ser Gly Gly Arg Ser Gly Arg 1100 1105 1110 Ser Gly Ser Phe Ile Tyr Gln Val Ser Thr His Glu Gln Ser Glu 1115 1120 1125 Ser Ala His Gly Arg Thr Arg Thr Ser Thr Gly Arg Arg Gln Gly 1130 1135 1140 Ser His His Glu Gln Ala Arg Asp Ser Ser Arg His Ser Ala Ser 1145 1150 1155 Gln Glu Gly Gln Asp Thr Ile Arg Ala His Pro Gly Ser Arg Arg 1160 1165 1170 Gly Gly Arg Gln Gly Ser His His Glu Gln Ser Val Asp Arg Ser 1175 1180 1185 Gly His Ser Gly Ser His His Ser His Thr Thr Ser Gln Gly Arg 1190 1195 1200 Ser Asp Ala Ser His Gly Gln Ser Gly Ser Arg Ser Ala Ser Arg 1205 1210 1215 Gln Thr Arg Lys Asp Lys Gln Ser Gly Asp Gly Ser Arg His Ser 1220 1225 1230 Gly Ser Arg His His Glu Ala Ala Ser Trp Ala Asp Ser Ser Arg 1235 1240 1245 His Ser Gln Val Gly Gln Glu Gln Ser Ser Gly Ser Arg Thr Ser 1250 1255 1260 Arg His Gln Gly Ser Ser Val Ser Gln Asp Ser Asp Ser Glu Arg 1265 1270 1275 His Ser Asp Asp Ser Glu Arg Leu Ser Gly Ser Ala Ser Arg Asn 1280 1285 1290 His His Gly Ser Ser Arg Glu Gln Ser Arg Asp Gly Ser Arg His 1295 1300 1305 Pro Gly Phe His Gln Glu Asp Arg Ala Ser His Gly His Ser Ala 1310 1315 1320 Asp Ser Ser Arg Gln Ser Gly Thr His His Thr Glu Ser Ser Ser 1325 1330 1335 His Gly Gln Ala Val Ser Ser His Glu Gln Ala Arg Ser Ser Pro 1340 1345 1350 Gly Glu Arg His Gly Ser Arg His Gln Gln Ser Ala Asp Ser Ser 1355 1360 1365 Arg His Ser Gly Ile Gly His Arg Gln Ala Ser Ser Ala Val Arg 1370 1375 1380 Asp Ser Gly His Arg Gly Ser Ser Gly Ser Gln Val Thr Asn Ser 1385 1390 1395 Glu Gly His Ser Glu Asp Ser Asp Thr Gln Ser Val Ser Ala His 1400 1405 1410 Gly Gln Ala Gly Pro His Gln Gln Ser His Lys Glu Ser Ala Arg 1415 1420 1425 Gly Gln Ser Gly Glu Ser Ser Gly Arg Ser Arg Ser Phe Leu Tyr 1430 1435 1440 Gln Val Ser Ser His Glu Gln Ser Glu Ser Thr His Gly Gln Thr 1445 1450 1455 Ala Pro Ser Thr Gly Gly Arg Gln Gly Ser Arg His Glu Gln Ala 1460 1465 1470 Arg Asn Ser Ser Arg His Ser Ala Ser Gln Asp Gly Gln Asp Thr 1475 1480 1485 Ile Arg Gly His Pro Gly Ser Ser Arg Gly Gly Arg Gln Gly Ser 1490 1495 1500 Tyr His Glu Gln Ser Val Asp Arg Ser Gly His Ser Gly Tyr His 1505 1510 1515 His Ser His Thr Thr Pro Gln Gly Arg Ser Asp Ala Ser His Gly 1520 1525 1530 Gln Ser Gly Pro Arg Ser Ala Ser Arg Gln Thr Arg Asn Glu Glu 1535 1540 1545 Gln Ser Gly Asp Gly Ser Arg His Ser Gly Ser Arg His His Glu 1550 1555 1560 Pro Ser Thr Arg Ala Gly Ser Ser Arg His Ser Gln Val Gly Gln 1565 1570 1575 Gly Glu Ser Ala Gly Ser Lys Thr Ser Arg Arg Gln Gly Ser Ser 1580 1585 1590 Val Ser Gln Asp Arg Asp Ser Glu Gly His Ser Glu Asp Ser Glu 1595 1600 1605 Arg Arg Ser Glu Ser Ala Ser Arg Asn His Tyr Gly Ser Ala Arg 1610 1615 1620 Glu Gln Ser Arg His Gly Ser Arg Asn Pro Arg Ser His Gln Glu 1625 1630 1635 Asp Arg Ala Ser His Gly His Ser Ala Glu Ser Ser Arg Gln Ser 1640 1645 1650 Gly Thr Arg His Ala Glu Thr Ser Ser Gly Gly Gln Ala Ala Ser 1655 1660 1665 Ser Gln Glu Gln Ala Arg Ser Ser Pro Gly Glu Arg His Gly Ser 1670 1675 1680 Arg His Gln Gln Ser Ala Asp Ser Ser Thr Asp Ser Gly Thr Gly 1685 1690 1695 Arg Arg Gln Asp Ser Ser Val Val Gly Asp Ser Gly Asn Arg Gly 1700 1705 1710 Ser Ser Gly Ser Gln Ala Ser Asp Ser Glu Gly His Ser Glu Glu 1715 1720 1725 Ser Asp Thr Gln Ser Val Ser Ala His Gly Gln Ala Gly Pro His 1730 1735 1740 Gln Gln Ser His Gln Glu Ser Thr Arg Gly Gln Ser Gly Glu Arg 1745 1750 1755 Ser Gly Arg Ser Gly Ser Phe Leu Tyr Gln Val Ser Thr His Glu 1760 1765 1770 Gln Ser Glu Ser Ala His Gly Arg Thr Gly Pro Ser Thr Gly Gly 1775 1780 1785 Arg Gln Arg Ser Arg His Glu Gln Ala Arg Asp Ser Ser Arg His 1790 1795 1800 Ser Ala Ser Gln Glu Gly Gln Asp Thr Ile Arg Gly His Pro Gly 1805 1810 1815 Ser Ser Arg Gly Gly Arg Gln Gly Ser His Tyr Glu Gln Ser Val 1820 1825 1830 Asp Ser Ser Gly His Ser Gly Ser His His Ser His Thr Thr Ser 1835 1840 1845 Gln Glu Arg Ser Asp Val Ser Arg Gly Gln Ser Gly Ser Arg Ser 1850 1855 1860 Val Ser Arg Gln Thr Arg Asn Glu Lys Gln Ser Gly Asp Gly Ser 1865 1870 1875 Arg His Ser Gly Ser Arg His His Glu Ala Ser Ser Arg Ala Asp 1880 1885 1890 Ser Ser Arg His Ser Gln Val Gly Gln Gly Gln Ser Ser Gly Pro 1895 1900 1905 Arg Thr Ser Arg Asn Gln Gly Ser Ser Val Ser Gln Asp Ser Asp 1910 1915 1920 Ser Gln Gly His Ser Glu Asp Ser Glu Arg Trp Ser Gly Ser Ala 1925 1930 1935 Ser Arg Asn His Leu Gly Ser Ala Trp Glu Gln Ser Arg Asp Gly 1940 1945 1950 Ser Arg His Pro Gly Ser His His Glu Asp Arg Ala Gly His Gly 1955 1960 1965 His Ser Ala Asp Ser Ser Arg Gln Ser Gly Thr Arg His Thr Glu 1970 1975 1980 Ser Ser Ser Arg Gly Gln Ala Ala Ser Ser His Glu Gln Ala Arg 1985 1990 1995 Ser Ser Ala Gly Glu Arg His Gly Ser His His Gln Leu Gln Ser 2000 2005 2010 Ala Asp Ser Ser Arg His Ser Gly Ile Gly His Gly Gln Ala Ser 2015 2020 2025 Ser Ala Val Arg Asp Ser Gly His Arg Gly Tyr Ser Gly Ser Gln 2030 2035 2040 Ala Ser Asp Ser Glu Gly His Ser Glu Asp Ser Asp Thr Gln Ser 2045 2050 2055 Val Ser Ala Gln Gly Lys Ala Gly Pro His Gln Gln Ser His Lys 2060 2065 2070 Glu Ser Ala Arg Gly Gln Ser Gly Glu Ser Ser Gly Arg Ser Gly 2075 2080 2085 Ser Phe Leu Tyr Gln Val Ser Thr His Glu Gln Ser Glu Ser Thr 2090 2095 2100 His Gly Gln Ser Ala Pro Ser Thr Gly Gly Arg Gln Gly Ser His 2105 2110 2115 Tyr Asp Gln Ala Gln Asp Ser Ser Arg His Ser Ala Ser Gln Glu 2120 2125 2130 Gly Gln Asp Thr Ile Arg Gly His Pro Gly Pro Ser Arg Gly Gly 2135 2140 2145 Arg Gln Gly Ser His Gln Glu Gln Ser Val Asp Arg Ser Gly His 2150 2155 2160 Ser Gly Ser His His Ser His Thr Thr Ser Gln Gly Arg Ser Asp 2165 2170 2175 Ala Ser Arg Gly Gln Ser Gly Ser Arg Ser Ala Ser Arg Lys Thr 2180 2185 2190 Tyr Asp Lys Glu Gln Ser Gly Asp Gly Ser Arg His Ser Gly Ser 2195 2200 2205 His His His Glu Ala Ser Ser Trp Ala Asp Ser Ser Arg His Ser 2210 2215 2220 Leu Val Gly Gln Gly Gln Ser Ser Gly Pro Arg Thr Ser Arg Pro 2225 2230 2235 Arg Gly Ser Ser Val Ser Gln Asp Ser Asp Ser Glu Gly His Ser 2240 2245 2250 Glu Asp Ser Glu Arg Arg Ser Gly Ser Ala Ser Arg Asn His His 2255 2260 2265 Gly Ser Ala Gln Glu Gln Ser Arg Asp Gly Ser Arg His Pro Arg 2270 2275 2280 Ser His His Glu Asp Arg Ala Gly His Gly His Ser Ala Glu Ser 2285 2290 2295 Ser Arg Gln Ser Gly Thr His His Ala Glu Asn Ser Ser Gly Gly 2300 2305 2310 Gln Ala Ala Ser Ser His Glu Gln Ala Arg Ser Ser Ala Gly Glu 2315 2320 2325 Arg His Gly Ser His His Gln Gln Ser Ala Asp Ser Ser Arg His 2330 2335 2340 Ser Gly Ile Gly His Gly Gln Ala Ser Ser Ala Val Arg Asp Ser 2345 2350 2355 Gly His Arg Gly Ser Ser Gly Ser Gln Ala Ser Asp Ser Glu Gly 2360 2365 2370 His Ser Glu Asp Ser Asp Thr Gln Ser Val Ser Ala His Gly Gln 2375 2380 2385 Ala Gly Pro His Gln Gln Ser His Gln Glu Ser Thr Arg Gly Arg 2390 2395 2400 Ser Ala Gly Arg Ser Gly Arg Ser Gly Ser Phe Leu Tyr Gln Val 2405 2410 2415 Ser Thr His Glu Gln Ser Glu Ser Ala His Gly Arg Thr Gly Thr 2420 2425 2430 Ser Thr Gly Gly Arg Gln Gly Ser His His Lys Gln Ala Arg Asp 2435 2440 2445 Ser Ser Arg His Ser Thr Ser Gln Glu Gly Gln Asp Thr Ile His 2450 2455 2460 Gly His Pro Gly Ser Ser Ser Gly Gly Arg Gln Gly Ser His Tyr 2465 2470 2475 Glu Gln Leu Val Asp Arg Ser Gly His Ser Gly Ser His His Ser 2480 2485 2490 His Thr Thr Ser Gln Gly Arg Ser Asp Ala Ser His Gly His Ser 2495 2500 2505 Gly Ser Arg Ser Ala Ser Arg Gln Thr Arg Asn Asp Glu Gln Ser 2510 2515 2520 Gly Asp Gly Ser Arg His Ser Gly Ser Arg His His Glu Ala Ser 2525 2530 2535 Ser Arg Ala Asp Ser Ser Gly His Ser Gln Val Gly Gln Gly Gln 2540 2545 2550 Ser Glu Gly Pro Arg Thr Ser Arg Asn Trp Gly Ser Ser Phe Ser 2555 2560 2565 Gln Asp Ser Asp Ser Gln Gly His Ser Glu Asp Ser Glu Arg Trp 2570 2575 2580 Ser Gly Ser Ala Ser Arg Asn His His Gly Ser Ala Gln Glu Gln 2585 2590 2595 Leu Arg Asp Gly Ser Arg His Pro Arg Ser His Gln Glu Asp Arg 2600 2605 2610 Ala Gly His Gly His Ser Ala Asp Ser Ser Arg Gln Ser Gly Thr 2615 2620 2625 Arg His Thr Gln Thr Ser Ser Gly Gly Gln Ala Ala Ser Ser His 2630 2635 2640 Glu Gln Ala Arg Ser Ser Ala Gly Glu Arg His Gly Ser His His 2645 2650 2655 Gln Gln Ser Ala Asp Ser Ser Arg His Ser Gly Ile Gly His Gly 2660 2665 2670 Gln Ala Ser Ser Ala Val Arg Asp Ser Gly His Arg Gly Tyr Ser 2675 2680 2685 Gly Ser Gln Ala Ser Asp Asn Glu Gly His Ser Glu Asp Ser Asp 2690 2695 2700 Thr Gln Ser Val Ser Ala His Gly Gln Ala Gly Ser His Gln Gln 2705 2710 2715 Ser His Gln Glu Ser Ala Arg Gly Arg Ser Gly Glu Thr Ser Gly 2720 2725 2730 His Ser Gly Ser Phe Leu Tyr Gln Val Ser Thr His Glu Gln Ser 2735 2740 2745 Glu Ser Ser His Gly Trp Thr Gly Pro Ser Thr Arg Gly Arg Gln 2750 2755 2760 Gly Ser Arg His Glu Gln Ala Gln Asp Ser Ser Arg His Ser Ala 2765 2770 2775 Ser Gln Asp Gly Gln Asp Thr Ile Arg Gly His Pro Gly Ser Ser 2780 2785 2790 Arg Gly Gly Arg Gln Gly Tyr His His Glu His Ser Val Asp Ser 2795 2800 2805 Ser Gly His Ser Gly Ser His His Ser His Thr Thr Ser Gln Gly 2810 2815 2820 Arg Ser Asp Ala Ser Arg Gly Gln Ser Gly Ser Arg Ser Ala Ser 2825 2830 2835 Arg Thr Thr Arg Asn Glu Glu Gln Ser Gly Asp Gly Ser Arg His 2840 2845 2850 Ser Gly Ser Arg His His Glu Ala Ser Thr His Ala Asp Ile Ser 2855 2860 2865 Arg His Ser Gln Ala Val Gln Gly Gln Ser Glu Gly Ser Arg Arg 2870 2875 2880 Ser Arg Arg Gln Gly Ser Ser Val Ser Gln Asp Ser Asp Ser Glu 2885 2890 2895 Gly His Ser Glu Asp Ser Glu Arg Trp Ser Gly Ser Ala Ser Arg 2900 2905 2910 Asn His His Gly Ser Ala Gln Glu Gln Leu Arg Asp Gly Ser Arg 2915 2920 2925 His Pro Arg Ser His Gln Glu Asp Arg Ala Gly His Gly His Ser 2930 2935 2940 Ala Asp Ser Ser Arg Gln Ser Gly Thr Arg His Thr Gln Thr Ser 2945 2950 2955 Ser Gly Gly Gln Ala Ala Ser Ser His Glu Gln Ala Arg Ser Ser 2960 2965 2970 Ala Gly Glu Arg His Gly Ser His His Gln Gln Ser Ala Asp Ser 2975 2980 2985 Ser Arg His Ser Gly Ile Gly His Gly Gln Ala Ser Ser Ala Val 2990 2995 3000 Arg Asp Ser Gly His Arg Gly Tyr Ser Gly Ser Gln Ala Ser Asp 3005 3010 3015 Asn Glu Gly His Ser Glu Asp Ser Asp Thr Gln Ser Val Ser Ala 3020 3025 3030 His Gly Gln Ala Gly Ser His Gln Gln Ser His Gln Glu Ser Ala 3035 3040 3045 Arg Gly Arg Ser Gly Glu Thr Ser Gly His Ser Gly Ser Phe Leu 3050 3055 3060 Tyr Gln Val Ser Thr His Glu Gln Ser Glu Ser Ser His Gly Trp 3065 3070 3075 Thr Gly Pro Ser Thr Arg Gly Arg Gln Gly Ser Arg His Glu Gln 3080 3085 3090 Ala Gln Asp Ser Ser Arg His Ser Ala Ser Gln Tyr Gly Gln Asp 3095 3100 3105 Thr Ile Arg Gly His Pro Gly Ser Ser Arg Gly Gly Arg Gln Gly 3110 3115 3120 Tyr His His Glu His Ser Val Asp Ser Ser Gly His Ser Gly Ser 3125 3130 3135 His His Ser His Thr Thr Ser Gln Gly Arg Ser Asp Ala Ser Arg 3140 3145 3150 Gly Gln Ser Gly Ser Arg Ser Ala Ser Arg Thr Thr Arg Asn Glu 3155 3160 3165 Glu Gln Ser Gly Asp Ser Ser Arg His Ser Val Ser Arg His His 3170 3175 3180 Glu Ala Ser Thr His Ala Asp Ile Ser Arg His Ser Gln Ala Val 3185 3190 3195 Gln Gly Gln Ser Glu Gly Ser Arg Arg Ser Arg Arg Gln Gly Ser 3200 3205 3210 Ser Val Ser Gln Asp Ser Asp Ser Glu Gly His Ser Glu Asp Ser 3215 3220 3225 Glu Arg Trp Ser Gly Ser Ala Ser Arg Asn His Arg Gly Ser Val 3230 3235 3240 Gln Glu Gln Ser Arg His Gly Ser Arg His Pro Arg Ser His His 3245 3250 3255 Glu Asp Arg Ala Gly His Gly His Ser Ala Asp Arg Ser Arg Gln 3260 3265 3270 Ser Gly Thr Arg His Ala Glu Thr Ser Ser Gly Gly Gln Ala Ala 3275 3280 3285 Ser Ser His Glu Gln Ala Arg Ser Ser Pro Gly Glu Arg His Gly 3290 3295 3300 Ser Arg His Gln Gln Ser Ala Asp Ser Ser Arg His Ser Gly Ile 3305 3310 3315 Pro Arg Gly Gln Ala Ser Ser Ala Val Arg Asp Ser Arg His Trp 3320 3325 3330 Gly Ser Ser Gly Ser Gln Ala Ser Asp Ser Glu Gly His Ser Glu 3335 3340 3345 Glu Ser Asp Thr Gln Ser Val Ser Gly His Gly Gln Ala Gly Pro 3350 3355 3360 His Gln Gln Ser His Gln Glu Ser Ala Arg Asp Arg Ser Gly Gly 3365 3370 3375 Arg Ser Gly Arg Ser Gly Ser Phe Leu Tyr Gln Val Ser Thr His 3380 3385 3390 Glu Gln Ser Glu Ser Ala His Gly Arg Thr Arg Thr Ser Thr Gly 3395 3400 3405 Arg Arg Gln Gly Ser His His Glu Gln Ala Arg Asp Ser Ser Arg 3410 3415 3420 His Ser Ala Ser Gln Glu Gly Gln Asp Thr Ile Arg Gly His Pro 3425 3430 3435 Gly Ser Ser Arg Arg Gly Arg Gln Gly Ser His Tyr Glu Gln Ser 3440 3445 3450 Val Asp Arg Ser Gly His Ser Gly Ser His His Ser His Thr Thr 3455 3460 3465 Ser Gln Gly Arg Ser Asp Ala Ser Arg Gly Gln Ser Gly Ser Arg 3470 3475 3480 Ser Ala Ser Arg Gln Thr Arg Asn Asp Glu Gln Ser Gly Asp Gly 3485 3490 3495 Ser Arg His Ser Trp Ser His His His Glu Ala Ser Thr Gln Ala 3500 3505 3510 Asp Ser Ser Arg His Ser Gln Ser Gly Gln Gly Gln Ser Ala Gly 3515 3520 3525 Pro Arg Thr Ser Arg Asn Gln Gly Ser Ser Val Ser Gln Asp Ser 3530 3535 3540 Asp Ser Gln Gly His Ser Glu Asp Ser Glu Arg Trp Ser Gly Ser 3545 3550 3555 Ala Ser Arg Asn His Arg Gly Ser Ala Gln Glu Gln Ser Arg Asp 3560 3565 3570 Gly Ser Arg His Pro Thr Ser His His Glu Asp Arg Ala Gly His 3575 3580 3585 Gly His Ser Ala Glu Ser Ser Arg Gln Ser Gly Thr His His Ala 3590 3595 3600 Glu Asn Ser Ser Gly Gly Gln Ala Ala Ser Ser His Glu Gln Ala 3605 3610 3615 Arg Ser Ser Ala Gly Glu Arg His Gly Ser His His Gln Gln Ser 3620 3625 3630 Ala Asp Ser Ser Arg His Ser Gly Ile Gly His Gly Gln Ala Ser 3635 3640 3645 Ser Ala Val Arg Asp Ser Gly His Arg Gly Ser Ser Gly Ser Gln 3650 3655 3660 Ala Ser Asp Ser Glu Gly His Ser Glu Asp Ser Asp Thr Gln Ser 3665 3670 3675 Val Ser Ala His Gly Gln Ala Gly Pro His Gln Gln Ser His Gln 3680 3685 3690 Glu Ser Thr Arg Gly Arg Ser Ala Gly Arg Ser Gly Arg Ser Gly 3695 3700 3705 Ser Phe Leu Tyr Gln Val Ser Thr His Glu Gln Ser Glu Ser Ala 3710 3715 3720 His Gly Arg Ala Gly Pro Ser Thr Gly Gly Arg Gln Gly Ser Arg 3725 3730 3735 His Glu Gln Ala Arg Asp Ser Ser Arg His Ser Ala Ser Gln Glu 3740 3745 3750 Gly Gln Asp Thr Ile Arg Gly His Pro Gly Ser Arg Arg Gly Gly 3755 3760 3765 Arg Gln Gly Ser Tyr His Glu Gln Ser Val Asp Arg Ser Gly His 3770 3775 3780 Ser Gly Ser His His Ser His Thr Thr Ser Gln Gly Arg Ser Asp 3785 3790 3795 Ala Ser His Gly Gln Ser Gly Ser Arg Ser Ala Ser Arg Glu Thr 3800 3805 3810 Arg Asn Glu Glu Gln Ser Gly Asp Gly Ser Arg His Ser Gly Ser 3815 3820 3825 Arg His His Glu Ala Ser Thr Gln Ala Asp Ser Ser Arg His Ser 3830 3835 3840 Gln Ser Gly Gln Gly Glu Ser Ala Gly Ser Arg Arg Ser Arg Arg 3845 3850 3855 Gln Gly Ser Ser Val Ser Gln Asp Ser Asp Ser Glu Ala Tyr Pro 3860 3865 3870 Glu Asp Ser Glu Arg Arg Ser Glu Ser Ala Ser Arg Asn His His 3875 3880 3885 Gly Ser Ser Arg Glu Gln Ser Arg Asp Gly Ser Arg His Pro Gly 3890 3895 3900 Ser Ser His Arg Asp Thr Ala Ser His Val Gln Ser Ser Pro Val 3905 3910 3915 Gln Ser Asp Ser Ser Thr Ala Lys Glu His Gly His Phe Ser Ser 3920 3925 3930 Leu Ser Gln Asp Ser Ala Tyr His Ser Gly Ile Gln Ser Arg Gly 3935 3940 3945 Ser Pro His Ser Ser Ser Ser Tyr His Tyr Gln Ser Glu Gly Thr 3950 3955 3960 Glu Arg Gln Lys Gly Gln Ser Gly Leu Val Trp Arg His Gly Ser 3965 3970 3975 Tyr Gly Ser Ala Asp Tyr Asp Tyr Gly Glu Ser Gly Phe Arg His 3980 3985 3990 Ser Gln His Gly Ser Val Ser Tyr Donkey Ser Donkey Pro Val Val Phe 3995 4000 4005 Lys Glu Arg Ser Asp Ile Cys Lys Ala Ser Ala Phe Gly Lys Asp 4010 4015 4020 His Pro Arg Tyr Tyr Ala Thr Tyr Ile Asn Lys Asp Pro Gly Leu 4025 4030 4035 Cys Gly His Ser Ser Asp Ile Ser Lys Gln Leu Gly Phe Ser Gln 4040 4045 4050 Ser Gln Arg Tyr Tyr Tyr Tyr Glu 4055 4060 <210> 12 <211> 4384 <212> PRT <213> Mus musculus <220> <221> MOD_RES <222> (1045)..(1045) <223> Any amino acid <220> <221> MOD_RES <222> (1048)..(1048) <223> Any amino acid <400> 12 Met Ser Ala Leu Leu Glu Ser Ile Thr Ser Met Ile Glu Ile Phe Gln 1 5 10 15 Gln Tyr Ser Thr Ser Asp Lys Glu Glu Glu Thr Leu Ser Lys Glu Glu 20 25 30 Leu Lys Glu Leu Leu Glu Gly Gln Leu Gln Ala Val Leu Lys Asn Pro 35 40 45 Asp Asp Gln Asp Ile Ala Glu Val Phe Met Gln Met Leu Asp Val Asp 50 55 60 His Asp Asp Lys Leu Asp Phe Ala Glu Tyr Leu Leu Leu Val Leu Lys 65 70 75 80 Leu Ala Lys Ala Tyr Tyr Glu Ala Ser Lys Asn Glu Ser Phe Gln Thr 85 90 95 His Gly Ser Asn Gly Arg Ser Lys Thr Asp Tyr Lys Gly Leu Glu Glu 100 105 110 Glu Gly Glu Glu Gly Asn Lys Gln Asn Leu Arg Arg Arg His Gly Gly 115 120 125 Thr Asp Gly Lys Arg Lys Ser Asp Arg Thr Arg Ser Pro Asn Gly Lys 130 135 140 Arg Gly Lys Arg Gln Glu Ser Arg Cys Arg Ser Glu Gly Lys Asp Lys 145 150 155 160 His Arg Arg Glu Pro Glu Lys His Arg His Gln Gln Asp Ser Lys Arg 165 170 175 Lys Gln Arg His Gly Ser Gly Ser Thr Glu Arg Lys Asp Asn Arg Asn 180 185 190 Lys Lys Asn Arg Gln Ser Lys Glu Arg Asn Tyr Asp Glu Ile Tyr Asp 195 200 205 Asn Gly Lys Tyr Asn Glu Asp Trp Glu Ala Ser Tyr Asn Asn Cys Tyr 210 215 220 Tyr Lys Thr Gln Asn Thr Thr Leu Asp Gln Arg Glu Gly Asn Arg Arg 225 230 235 240 Pro Arg Ala Asp Ser Gln Lys Glu Pro Gln Ser Ser His Gly Gln Ala 245 250 255 Asp Asn Ser Asp Ser Glu Gly Gly Arg Gln Gln Ser His Ser Lys Pro 260 265 270 Ser Pro Val Arg Ala Asp Gln Arg Arg Ser Arg Ala Gly Gln Ala Gly 275 280 285 Ser Ser Lys Val Ser Ala Arg Ser Gly Ser Gly Gly Arg Gly Gln Ser 290 295 300 Pro Asp Gly Ser Gly Arg Ser Ser Asn Arg Arg Asp Arg Pro Arg Gln 305 310 315 320 Pro Ser Pro Ser Gln Ser Ser Asp Ser Gln Val His Ser Gly Val Gln 325 330 335 Val Glu Gly Arg Arg Gly Gln Ser Ser Ser Ala Asn Arg Arg Ala Gly 340 345 350 Ser Ser Ser Gly Ser Gly Val Gln Gly Ala Ser Ala Gly Gly Leu Ala 355 360 365 Ala Asp Ala Ser Arg Arg Ser Gly Ala Arg Gln Gly Gln Ala Ser Ala 370 375 380 Gln Gly Arg Ala Gly Ser Gln Gly Gln Ala Gln Gly Arg Val Gly Ser 385 390 395 400 Ser Ala Asp Arg Gln Gly Arg Arg Gly Val Ser Glu Ser Gln Ala Ser 405 410 415 Asp Ser Glu Gly His Ser Asp Phe Ser Glu Gly Gln Ala Val Gly Ala 420 425 430 His Arg Gln Ser Gly Ala Gly Gln Arg His Glu Gln Arg Ser Ser Arg 435 440 445 Gly Gln His Gly Ser Arg Tyr Tyr Tyr Glu Gln Glu His Ser Glu Glu 450 455 460 Glu Ser Asp Ser Gln His Gln His Gly His Gln His Glu Gln Gln Arg 465 470 475 480 Gly His Gln His Gln His Glu His Glu Gln Pro Glu Ser Gly His Arg 485 490 495 Gln Gln Gln Ser Ser Gly Arg Gly His Gln Gly Ala His Gln Glu Gln 500 505 510 Gly Arg Asp Ser Ala Arg Ser Arg Gly Ser Asn Gln Gly His Ser Ser 515 520 525 Ser Arg His Gln Ala Asp Ser Pro Arg Val Ser Ala Arg Ser Gly Ser 530 535 540 Gly Gly Arg Gly Gln Ser Pro Asp Ala Ser Gly Arg Ser Ser Asn Arg 545 550 555 560 Arg Asp Arg Pro Arg Gln Pro Ser Pro Ser Gln Ser Ser Asp Ser Gln 565 570 575 Val His Ser Gly Val Gln Val Glu Gly Arg Arg Gly Gln Ser Ser Ser 580 585 590 Ala Asn Arg Arg Ala Gly Ser Ser Ser Gly Ser Gly Val Gln Gly Ala 595 600 605 Ser Ala Gly Gly Leu Ala Ala Asp Ala Ser Arg Arg Ser Gly Ala Leu 610 615 620 Gln Gly Gln Ala Ser Ala Gln Gly Arg Ala Gly Ser Gln Gly Gln Ala 625 630 635 640 Gln Gly Arg Val Gly Ser Ser Ala Asp Arg Gln Gly Arg Arg Gly Val 645 650 655 Ser Glu Ser Gln Ala Ser Asp Ser Glu Gly His Ser Asp Phe Ser Glu 660 665 670 Gly Gln Ala Val Gly Ala His Arg Gln Ser Gly Ala Gly Gln Arg His 675 680 685 Glu Gln Arg Ser Ser Arg Gly Gln His Gly Ser Gly Tyr Tyr Tyr Glu 690 695 700 Gln Glu His Ser Glu Glu Glu Ser Asp Ser Gln His Gln His Gly His 705 710 715 720 Gln His Glu Gln Gln Arg Gly His Gln His Gln His Gln His Gln His 725 730 735 Glu His Glu Gln Pro Glu Ser Gly His Arg Gln Gln Gln Ser Ser Gly 740 745 750 Arg Gly His Gln Gly Ala His Gln Glu Gln Gly Arg Asp Ser Ala Arg 755 760 765 Ser Arg Gly Ser Asn Gln Gly His Ser Ser Ser Arg His Gln Ala Asp 770 775 780 Ser Pro Arg Val Ser Ala Arg Ser Gly Ser Gly Gly Arg Gly Gln Ser 785 790 795 800 Pro Asp Ala Ser Gly Arg Ser Ser Asn Arg Arg Asp Arg Pro Arg Gln 805 810 815 Pro Ser Pro Ser Gln Ser Ser Asp Ser Gln Val His Ser Gly Val Gln 820 825 830 Val Glu Gly Arg Arg Gly Gln Ser Ser Ser Ala Asn Arg Arg Ala Gly 835 840 845 Ser Ser Ser Gly Ser Gly Val Gln Gly Thr Ser Ala Gly Gly Leu Ala 850 855 860 Ala Asp Ala Ser Arg Arg Ser Gly Ala Arg Gln Gly Gln Ala Ser Ala 865 870 875 880 Gln Gly Arg Ala Gly Ser Gln Gly Gln Ala Gln Gly Arg Val Gly Ser 885 890 895 Ser Ala Asp Arg Gln Gly Arg Arg Gly Val Ser Glu Ser Gln Ala Ser 900 905 910 Asp Ser Glu Gly His Ser Asp Phe Ser Glu Gly Gln Ala Val Gly Ala 915 920 925 His Arg Gln Ser Gly Ala Gly Gln Arg His Glu Gln Arg Ser Ser Arg 930 935 940 Gly Gln His Gly Ser Gly Tyr Tyr Tyr Glu Gln Glu His Ser Glu Glu 945 950 955 960 Glu Ser Asp Ser Gln His Gln His Gly His Gln His Glu Gln Gln Arg 965 970 975 Gly His Gln His Gln His Gln His Gln His Glu His Glu Gln Pro Glu 980 985 990 Ser Gly His Arg Gln Gln Gln Ser Ser Gly Arg Gly Asn Gln Gly Ala 995 1000 1005 His Gln Lys Gln Gly Arg Asp Ser Ala Arg Ser Arg Gly Ser Asn 1010 1015 1020 Gln Gly His Ser Ser Ser Arg His Gln Ala Asp Ser Pro Arg Val 1025 1030 1035 Ser Ala Arg Ser Gly Ser Xaa Gly Arg Xaa Gln Ser Pro Asp Ala 1040 1045 1050 Ser Gly Arg Asn Ser Thr Lys Arg Asp Arg Pro Arg Gln Pro Ser 1055 1060 1065 Pro Ser Gln Ser Ser Asp Ser His Val His Ser Gly Lys Ala Gly 1070 1075 1080 Ser Ile Ser Gly Ser Gly Val Gln Gly Ala Ser Ala Gly Gly Leu 1085 1090 1095 Ala Ala Asp Ala Ser Arg Arg Ser Gly Ala Arg Gln Gly Gln Ala 1100 1105 1110 Ser Ala Gln Gly Arg Ala Gly Ser Gln Gly Gln Ala Gln Gly Arg 1115 1120 1125 Val Gly Ser Ser Ala Asp Arg Gln Gly Arg Arg Gly Val Ser Glu 1130 1135 1140 Ser Gln Ala Ser Asp Ser Glu Gly His Ser Asp Phe Ser Glu Gly 1145 1150 1155 Gln Ala Val Gly Ala His Arg Gln Ser Gly Ala Gly Gln Arg His 1160 1165 1170 Glu Gln Arg Ser Ser Arg Gly Gln His Gly Ser Arg Tyr Tyr Tyr 1175 1180 1185 Glu Gln Glu His Ser Glu Glu Glu Ser Asp Ser Gln His Gln His 1190 1195 1200 Gly His Gln His Glu Gln Gln Arg Gly His Gln His Gln His Glu 1205 1210 1215 His Glu Gln Pro Glu Ser Gly His Arg Gln Gln Gln Ser Ser Gly 1220 1225 1230 Arg Gly His Gln Gly Ala His Gln Glu Gln Gly Arg Asp Ser Ala 1235 1240 1245 Arg Ser Arg Gly Ser Asn Gln Gly His Ser Ser Ser Arg His Gln 1250 1255 1260 Ala Asp Ser Pro Arg Val Ser Ala Arg Ser Gly Ser Gly Gly Arg 1265 1270 1275 Gly Gln Ser Pro Asp Ala Ser Gly Arg Ser Ser Asn Arg Arg Asp 1280 1285 1290 Arg Pro Arg Gln Pro Ser Pro Ser Gln Ser Ser Asp Ser Gln Val 1295 1300 1305 His Ser Gly Val Gln Val Glu Gly Arg Arg Gly Gln Ser Ser Ser 1310 1315 1320 Ala Asn Arg Arg Ala Gly Ser Ser Ser Gly Ser Gly Val Gln Gly 1325 1330 1335 Ala Ser Ala Gly Gly Leu Ala Ala Asp Ala Ser Arg Arg Ser Gly 1340 1345 1350 Ala Arg Gln Gly Gln Ala Ser Ala Gln Gly Arg Ala Gly Ser Gln 1355 1360 1365 Gly Gln Ala Gln Gly Arg Ile Gly Ser Ser Ala Asp Arg Gln Gly 1370 1375 1380 Arg Arg Gly Val Ser Glu Ser Gln Ala Ser Asp Ser Glu Gly His 1385 1390 1395 Ser Asp Phe Ser Glu Gly Gln Ala Val Gly Ala His Arg Gln Ser 1400 1405 1410 Glu Ala Gly Gln Arg His Glu Gln Arg Ser Ser Arg Gly Gln His 1415 1420 1425 Gly Ser Gly Tyr Tyr Tyr Glu Gln Glu His Ser Glu Glu Glu Ser 1430 1435 1440 Asp Ser Gln His Gln His Gly His Gln His Glu Gln Gln Arg Gly 1445 1450 1455 Thr Gln His Gln His Glu His Gln Gln Pro Glu Ser Gly His Arg 1460 1465 1470 Gln Gln Gln Ser Ser Gly Arg Gly His Gln Gly Thr His Gln Glu 1475 1480 1485 Gln Gly Arg Asp Ser Ala Arg Ser Arg Gly Ser Asn Gln Gly His 1490 1495 1500 Ser Ser Ser Arg His Gln Ala Asp Ser Pro Arg Val Ser Ala Arg 1505 1510 1515 Ser Gly Ser Gly Gly Arg Gly Gln Ser Pro Asp Ala Ser Gly Arg 1520 1525 1530 Ser Ser Asn Arg Arg Asp Arg Pro Arg Gln Pro Ser Pro Ser Gln 1535 1540 1545 Ser Ser Asp Ser Gln Val His Ser Gly Val Gln Val Glu Gly Arg 1550 1555 1560 Arg Arg Gln Ser Ser Ser Ala Asn Arg Arg Ala Gly Ser Ser Ser 1565 1570 1575 Gly Ser Gly Val Gln Gly Ala Ser Ala Gly Gly Leu Ala Ala Asp 1580 1585 1590 Ala Ser Arg Arg Ser Gly Ala Arg Gln Gly Gln Ala Ser Ala Gln 1595 1600 1605 Gly Arg Ala Gly Ser Gln Gly Gln Ala Gln Gly Arg Val Gly Ser 1610 1615 1620 Ser Ala Asp Arg Gln Gly Arg Arg Gly Val Ser Glu Ser Gln Ala 1625 1630 1635 Ser Asp Ser Glu Gly His Ser Asp Phe Ser Glu Gly Gln Ala Val 1640 1645 1650 Gly Ala His Arg Gln Ser Gly Ala Gly Gln Arg His Glu Gln Arg 1655 1660 1665 Ser Ser Arg Gly Gln His Gly Ser Gly Phe Tyr Pro Val Tyr Tyr 1670 1675 1680 Tyr Tyr Glu Gln Glu His Ser Glu Glu Glu Ser Asp Ser Gln His 1685 1690 1695 Gln His Gly His Gln His Glu Gln Gln Arg Gly His Gln His Gln 1700 1705 1710 His Gln His Glu His Glu Gln Pro Glu Ser Gly His Arg Gln Gln 1715 1720 1725 Gln Ser Ser Gly Arg Gly His Gln Gly Ala His Gln Glu Gln Gly 1730 1735 1740 Arg Asp Ser Ala Arg Ser Arg Gly Ser Asn Gln Gly His Ser Ser 1745 1750 1755 Ser Arg His Gln Ala Asp Ser Pro Arg Val Ser Ala Arg Ser Gly 1760 1765 1770 Ser Gly Gly Arg Gly Gln Ser Pro Asp Ala Ser Gly Arg Ser Ser 1775 1780 1785 Asn Arg Arg Asp Arg Pro Arg Gln Pro Ser Pro Ser Gln Ser Ser 1790 1795 1800 Asp Ser Gln Val His Ser Gly Val Gln Val Glu Gly Arg Arg Gly 1805 1810 1815 Gln Ser Ser Ser Ala Asn Arg Arg Ala Gly Ser Ser Ser Gly Ser 1820 1825 1830 Gly Val Gln Gly Ala Ser Ala Gly Gly Leu Ala Ala Asp Ala Ser 1835 1840 1845 Arg Arg Ser Gly Ala Leu Gln Gly Gln Ala Ser Ala Gln Gly Arg 1850 1855 1860 Ala Gly Ser Gln Gly Gln Ala Gln Gly Arg Val Gly Ser Ser Ala 1865 1870 1875 Asp Arg Gln Gly Arg Arg Gly Val Ser Gly Ser Gln Ala Ser Asp 1880 1885 1890 Ser Glu Gly His Ser Asp Phe Ser Glu Gly Gln Ala Val Gly Ala 1895 1900 1905 His Arg Gln Ser Gly Ala Gly Gln Arg His Glu Gln Arg Ser Ser 1910 1915 1920 Arg Gly Gln His Gly Ser Gly Tyr Tyr Tyr Glu Gln Glu His Ser 1925 1930 1935 Glu Glu Glu Ser Asp Ser Gln His Gln His Gly His Gln His Glu 1940 1945 1950 Gln Gln Arg Gly His Gln His Gln His Gln His Gln His Glu His 1955 1960 1965 Glu Gln Pro Glu Ser Gly His Arg Gln Gln Gln Phe Ser Gly Arg 1970 1975 1980 Gly His Gln Gly Ala His Gln Glu Gln Gly Arg Asp Ser Ala Arg 1985 1990 1995 Ser Arg Gly Ser Asn Gln Gly His Ser Ser Ser Arg His Gln Ala 2000 2005 2010 Asp Ser Pro Arg Val Ser Ala Arg Ser Gly Ser Gly Gly Arg Gly 2015 2020 2025 Gln Ser Pro Asp Ala Ser Gly Arg Ser Ser Asn Arg Arg Asp Arg 2030 2035 2040 Pro Arg Gln Pro Ser Pro Ser Gln Ser Ser Asp Ser Gln Val His 2045 2050 2055 Ser Gly Val Gln Val Glu Gly Arg Arg Gly Gln Ser Ser Ser Ala 2060 2065 2070 Asn Arg Arg Ala Gly Ser Ser Ser Gly Ser Gly Val Gln Gly Ala 2075 2080 2085 Ser Ala Gly Gly Leu Ala Ala Asp Ala Ser Arg Arg Ser Gly Ala 2090 2095 2100 Arg Gln Gly Gln Ala Ser Ala Gln Gly Arg Ala Gly Ser Gln Gly 2105 2110 2115 Gln Ala Gln Gly Arg Val Gly Ser Ser Ala Asp Arg Gln Gly Arg 2120 2125 2130 Arg Gly Val Ser Glu Ser Gln Ala Ser Asp Ser Glu Gly His Ser 2135 2140 2145 Asp Phe Ser Glu Gly Gln Ala Val Gly Ala His Arg Gln Ser Gly 2150 2155 2160 Ala Gly Gln Arg His Glu Gln Arg Ser Ser Arg Gly Gln His Gly 2165 2170 2175 Ser Gly Phe Tyr Pro Val Tyr Tyr Tyr Tyr Glu Gln Glu His Ser 2180 2185 2190 Glu Glu Glu Ser Asp Ser Gln His Gln His Gly His Gln His Glu 2195 2200 2205 Gln Gln Arg Gly His Gln His Gln His Gln His Gln His Glu His 2210 2215 2220 Glu Gln Pro Glu Ser Gly His Arg Gln Gln Gln Ser Ser Gly Arg 2225 2230 2235 Gly His Gln Gly Ala His Gln Glu Gln Gly Arg Asp Ser Ala Arg 2240 2245 2250 Ser Arg Gly Ser Asn Gln Gly His Ser Ser Ser Arg His Gln Ala 2255 2260 2265 Asp Ser Pro Arg Val Ser Ala Arg Ser Gly Ser Gly Gly Arg Gly 2270 2275 2280 Gln Ser Pro Asp Ala Ser Gly Arg Ser Ser Asn Arg Arg Asp Arg 2285 2290 2295 Pro Arg Gln Pro Ser Pro Ser Gln Ser Ser Asp Ser Gln Val His 2300 2305 2310 Ser Gly Val Gln Val Glu Gly Arg Arg Gly Gln Ser Ser Ser Ala 2315 2320 2325 Asn Arg Arg Ala Gly Ser Ser Ser Gly Ser Gly Val Gln Gly Ala 2330 2335 2340 Ser Ala Gly Gly Leu Ala Ala Asp Ala Ser Arg Arg Ser Gly Ala 2345 2350 2355 Arg Gln Gly Gln Ala Ser Ala Gln Gly Arg Ala Gly Ser Gln Gly 2360 2365 2370 Gln Ala Gln Gly Arg Val Gly Ser Ser Ala Asp Arg Gln Gly Arg 2375 2380 2385 Arg Gly Val Ser Glu Ser Gln Ala Ser Asp Ser Glu Gly His Ser 2390 2395 2400 Asp Phe Ser Glu Gly Gln Ala Val Gly Ala His Arg Gln Ser Gly 2405 2410 2415 Ala Gly Gln Arg His Glu Gln Arg Ser Ser Arg Gly Gln His Gly 2420 2425 2430 Ser Gly Tyr Tyr Tyr Glu Gln Glu His Ser Glu Glu Glu Ser Asp 2435 2440 2445 Ser Gln His Gln His Gly His Gln His Glu Gln Gln Arg Gly His 2450 2455 2460 Gln His Gln His Gln His Gln His Glu His Glu Gln Pro Glu Ser 2465 2470 2475 Gly His Arg Gln Gln Gln Ser Ser Gly Arg Gly His Gln Gly Ala 2480 2485 2490 His Gln Glu Gln Gly Arg Asp Ser Ala Arg Ser Arg Gly Ser Asn 2495 2500 2505 Gln Gly His Ser Ser Ser Arg His Gln Ala Asp Ser Pro Arg Val 2510 2515 2520 Ser Ala Arg Ser Gly Ser Gly Gly Arg Gly Gln Ser Pro Asp Ala 2525 2530 2535 Ser Gly Arg Ser Ser Asn Arg Arg Asp Arg Pro Arg Gln Pro Ser 2540 2545 2550 Pro Ser Gln Ser Ser Asp Ser Gln Val His Ser Gly Val Gln Val 2555 2560 2565 Glu Gly Arg Arg Gly Gln Ser Ser Ser Ala Asn Arg Arg Ala Gly 2570 2575 2580 Ser Ser Ser Gly Ser Gly Val Gln Gly Ala Ser Ala Gly Gly Leu 2585 2590 2595 Ala Ala Asp Ala Ser Arg Arg Ser Gly Ala Arg Gln Gly Gln Ala 2600 2605 2610 Ser Ala Gln Gly Arg Ala Gly Ser Gln Gly Gln Ala Gln Gly Arg 2615 2620 2625 Val Gly Ser Ser Ala Asp Arg Gln Gly Arg Arg Gly Val Ser Glu 2630 2635 2640 Ser Gln Ala Ser Asp Ser Glu Gly His Ser Asp Phe Ser Glu Gly 2645 2650 2655 Gln Ala Val Gly Ala His Arg Gln Ser Gly Ala Gly Gln Arg His 2660 2665 2670 Glu Gln Arg Ser Ser Arg Ser Gln His Gly Ser Gly Tyr Tyr Tyr 2675 2680 2685 Glu Gln Glu His Ser Glu Glu Glu Ser Asp Ser Gln His Gln His 2690 2695 2700 Ser His Gln His Glu Gln Gln Arg Gly His Gln His Gln His Gln 2705 2710 2715 His Gln His Glu His Glu Gln Pro Glu Ser Gly His Arg Gln Gln 2720 2725 2730 Gln Ser Ser Gly Arg Gly Asn Gln Gly Ala His Gln Glu Gln Gly 2735 2740 2745 Arg Asp Ser Ala Arg Ser Arg Gly Ser Asn Gln Gly His Ser Ser 2750 2755 2760 Ser Arg His Gln Ala Asp Ser Pro Arg Val Ser Ala Arg Ser Gly 2765 2770 2775 Ser Gly Gly Arg Gly Gln Ser Pro Asp Ala Ser Gly Arg Ser Ser 2780 2785 2790 Asn Arg Arg Asp Arg Pro Arg Gln Pro Ser Pro Ser Gln Ser Ser 2795 2800 2805 Asp Ser His Val His Ser Gly Val Gln Val Glu Gly Arg Arg Gly 2810 2815 2820 Gln Ser Ser Ser Ala Asn Arg Arg Ala Gly Ser Ser Ser Gly Ser 2825 2830 2835 Gly Val Gln Gly Ala Ser Ala Gly Gly Leu Ala Ala Asp Ala Ser 2840 2845 2850 Arg Arg Ser Gly Ala Arg Gln Gly Gln Ala Ser Ala Gln Gly Arg 2855 2860 2865 Ala Gly Ser Gln Gly Gln Ala Gln Gly Arg Val Gly Ser Ser Ala 2870 2875 2880 Asp Arg Gln Gly Arg Arg Gly Val Ser Glu Ser Gln Ala Ser Asp 2885 2890 2895 Ser Glu Gly His Ser Asp Phe Ser Glu Gly Gln Ala Val Gly Ala 2900 2905 2910 His Arg Gln Ser Gly Ala Gly Gln Arg His Glu Gln Arg Ser Ser 2915 2920 2925 Arg Gly Gln His Gly Ser Gly Phe Tyr Pro Val Tyr Tyr Tyr Tyr 2930 2935 2940 Glu Gln Glu His Ser Glu Glu Glu Ser Asp Ser Gln His Gln His 2945 2950 2955 Gly His Gln His Glu Gln Gln Arg Gly His Gln His Gln His Gln 2960 2965 2970 His Gln His Glu His Glu Gln Pro Glu Ser Gly His Arg Gln Gln 2975 2980 2985 Gln Ser Ser Gly Arg Gly His Gln Gly Ala His Gln Glu Gln Gly 2990 2995 3000 Arg Asp Ser Ala Arg Ser Arg Gly Ser Asn Gln Gly His Ser Ser 3005 3010 3015 Ser Arg His Gln Ala Asp Ser Pro Arg Val Ser Ala Arg Ser Gly 3020 3025 3030 Ser Gly Gly Arg Gly Gln Ser Pro Asp Ala Ser Gly Arg Ser Ser 3035 3040 3045 Asn Arg Arg Asp Arg Ser Arg Gln Pro Ser Pro Ser Gln Ser Ser 3050 3055 3060 Asp Ser Gln Val His Ser Gly Val Gln Val Glu Gly Arg Arg Gly 3065 3070 3075 Gln Ser Ser Ser Ala Asn Arg Arg Ala Gly Ser Ser Ser Gly Ser 3080 3085 3090 Gly Val Gln Gly Ala Ser Ala Gly Gly Leu Ala Ala Asp Ala Ser 3095 3100 3105 Arg Arg Ser Gly Ala Arg Gln Gly Gln Ala Ser Ala Gln Gly Arg 3110 3115 3120 Ala Gly Ser Gln Gly Gln Ala Gln Gly Arg Val Gly Ser Ser Ala 3125 3130 3135 Asp Arg Gln Gly Arg Arg Gly Val Ser Glu Ser Gln Ala Ser Asp 3140 3145 3150 Ser Glu Gly His Ser Asp Phe Ser Glu Gly Gln Ala Val Gly Ala 3155 3160 3165 His Arg Gln Ser Gly Ala Gly Gln Arg His Glu Gln Arg Ser Ser 3170 3175 3180 Arg Gly Gln His Gly Ser Gly Phe Tyr Pro Val Tyr Tyr Tyr Tyr 3185 3190 3195 Glu Gln Glu His Ser Glu Glu Glu Ser Asp Ser Gln His Gln His 3200 3205 3210 Gly His Gln His Glu Gln Gln Arg Gly His Gln His Gln His Gln 3215 3220 3225 His Glu His Glu Gln Pro Glu Ser Gly His Arg Gln Gln Gln Ser 3230 3235 3240 Ser Gly Arg Gly His Gln Gly Ala His Gln Glu Gln Gly Arg Asp 3245 3250 3255 Ser Ala Arg Ser Arg Gly Ser Asn Gln Gly His Ser Ser Ser Arg 3260 3265 3270 His Gln Ala Asp Ser Pro Arg Val Ser Ala Arg Ser Gly Ser Gly 3275 3280 3285 Gly Arg Gly Gln Ser Pro Asp Ala Ser Gly Arg Ser Ser Asn Arg 3290 3295 3300 Arg Asp Arg Pro Arg Gln Pro Ser Pro Ser Gln Ser Ser Asp Ser 3305 3310 3315 Gln Val His Ser Gly Val Gln Val Glu Gly Arg Arg Gly Gln Ser 3320 3325 3330 Ser Ser Ala Asn Arg Arg Ala Gly Ser Ser Ser Gly Ser Gly Val 3335 3340 3345 Gln Gly Ala Ser Ala Gly Gly Leu Ala Ala Asp Ala Ser Arg Arg 3350 3355 3360 Ser Gly Ala Leu Gln Gly Gln Ala Ser Ala Gln Gly Arg Ala Gly 3365 3370 3375 Ser Gln Gly Gln Ala Gln Gly Arg Val Gly Ser Ser Ala Asp Arg 3380 3385 3390 Gln Gly Arg Arg Gly Val Ser Glu Ser Gln Ala Ser Asp Ser Glu 3395 3400 3405 Gly His Ser Asp Phe Ser Glu Gly Gln Ala Val Gly Ala His Arg 3410 3415 3420 Gln Ser Gly Ala Gly Gln Arg His Glu Gln Arg Ser Ser Arg Gly 3425 3430 3435 Gln His Gly Ser Gly Tyr Tyr Tyr Glu Gln Glu His Ser Glu Glu 3440 3445 3450 Glu Ser Asp Ser Gln His Gln His Gly His Gln His Glu Gln Gln 3455 3460 3465 Arg Gly His Gln His Gln His Gln His Gln His Glu His Glu Gln 3470 3475 3480 Pro Glu Ser Gly His Arg Gln Gln Gln Ser Ser Gly Arg Gly His 3485 3490 3495 Gln Gly Ala His Gln Glu Gln Gly Arg Asp Ser Ala Arg Ser Arg 3500 3505 3510 Gly Ser Asn Gln Gly His Ser Ser Ser Arg His Gln Ala Asp Ser 3515 3520 3525 Pro Arg Val Ser Ala Arg Ser Gly Ser Gly Gly Arg Gly Gln Ser 3530 3535 3540 Pro Asp Ala Ser Gly Arg Ser Ser Asn Arg Arg Asp Arg Pro Arg 3545 3550 3555 Gln Pro Ser Pro Ser Gln Ser Ser Asp Ser Gln Val His Ser Gly 3560 3565 3570 Val Gln Val Glu Gly Arg Arg Gly Gln Ser Ser Ser Ala Asn Arg 3575 3580 3585 Arg Ala Gly Ser Ser Ser Gly Ser Gly Val Gln Gly Ala Ser Ala 3590 3595 3600 Gly Gly Leu Ala Ala Asp Ala Ser Arg Arg Ser Gly Ala Arg Gln 3605 3610 3615 Gly Gln Ala Ser Ala Gln Gly Arg Ala Gly Ser Gln Gly Gln Ala 3620 3625 3630 Gln Gly Arg Val Gly Ser Ser Ala Asp Arg Gln Gly Arg Arg Gly 3635 3640 3645 Val Ser Gly Ser Gln Ala Ser Asp Ser Glu Gly His Ser Asp Phe 3650 3655 3660 Ser Glu Gly Gln Ala Val Gly Ala His Arg Gln Ser Gly Ala Gly 3665 3670 3675 Gln Arg His Glu Gln Arg Ser Ser Arg Gly Gln His Gly Ser Gly 3680 3685 3690 Tyr Tyr Tyr Glu Gln Glu His Ser Glu Glu Glu Ser Asp Ser Gln 3695 3700 3705 His Gln His Gly His Gln His Glu Gln Gln Arg Gly His Gln His 3710 3715 3720 Gln His Gln His Gln His Glu His Glu Gln Pro Glu Ser Gly His 3725 3730 3735 Arg Gln Gln Gln Phe Ser Gly Arg Gly His Gln Gly Ala His Gln 3740 3745 3750 Glu Gln Gly Arg Asp Ser Ala Arg Ser Arg Gly Ser Asn Gln Gly 3755 3760 3765 His Ser Ser Ser Arg His Gln Ala Asp Ser Pro Arg Val Ser Ala 3770 3775 3780 Arg Ser Gly Ser Gly Gly Arg Gly Gln Ser Pro Asp Ala Ser Gly 3785 3790 3795 Arg Ser Ser Asn Arg Arg Asp Arg Pro Arg Gln Pro Ser Pro Ser 3800 3805 3810 Gln Ser Ser Asp Ser Gln Val His Ser Gly Val Gln Val Glu Gly 3815 3820 3825 Arg Arg Gly Gln Ser Ser Ser Ala Asn Arg Arg Ala Gly Ser Ser 3830 3835 3840 Ser Ser Ser Gly Val Gln Gly Ala Ser Ala Gly Gly Leu Ala Ala 3845 3850 3855 Asp Ala Ser Arg Arg Ser Gly Ala Arg Gln Gly Gln Ala Ser Ala 3860 3865 3870 Gln Gly Arg Ala Gly Ser Gln Gly Gln Ala Gln Gly Arg Val Gly 3875 3880 3885 Ser Ser Ala Asp Arg Gln Gly Arg Arg Gly Val Ser Glu Ser Gln 3890 3895 3900 Ala Ser Asp Ser Glu Gly His Ser Asp Phe Ser Glu Gly Gln Ala 3905 3910 3915 Val Gly Ala His Arg Gln Ser Gly Ala Gly Gln Arg His Glu Gln 3920 3925 3930 Arg Ser Ser Arg Gly Gln His Gly Ser Gly Phe Tyr Pro Val Tyr 3935 3940 3945 Tyr Tyr Tyr Glu Gln Glu His Ser Glu Glu Glu Ser Asp Ser Gln 3950 3955 3960 His Gln His Gly His Gln His Glu Gln Gln Arg Gly His Gln His 3965 3970 3975 Gln His Gln His Gln His Glu His Glu Gln Pro Glu Ser Gly His 3980 3985 3990 Arg Gln Gln Gln Phe Ser Gly Arg Gly His Gln Gly Ala His Gln 3995 4000 4005 Glu Gln Gly Arg Asp Ser Ala Arg Ser Arg Gly Ser Asn Gln Gly 4010 4015 4020 His Ser Ser Ser Arg His Gln Ala Asp Ser Pro Arg Val Ser Ala 4025 4030 4035 Arg Ser Gly Ser Gly Gly Arg Gly Gln Ser Pro Asp Ala Ser Gly 4040 4045 4050 Arg Ser Ser Asn Arg Arg Asp Arg Pro Arg Gln Pro Ser Pro Ser 4055 4060 4065 Gln Ser Ser Asp Ser Gln Val His Ser Gly Val Gln Val Glu Gly 4070 4075 4080 Arg Arg Gly Gln Ser Ser Ser Ala Asn Arg Arg Ala Gly Ser Ser 4085 4090 4095 Ser Gly Ser Gly Val Gln Gly Ala Ser Ala Gly Gly Leu Ala Ala 4100 4105 4110 Asp Ala Ser Arg Arg Ser Gly Ala Arg Gln Gly Gln Ala Ser Ala 4115 4120 4125 Gln Gly Arg Ala Gly Ser Gln Gly Gln Ala Gln Gly Arg Val Gly 4130 4135 4140 Ser Ser Ala Asp Arg Gln Gly Arg Arg Gly Val Ser Glu Ser Gln 4145 4150 4155 Ala Ser Asp Ser Glu Gly His Ser Asp Phe Ser Glu Gly Gln Ala 4160 4165 4170 Val Gly Ala His Arg Gln Ser Gly Ala Gly Gln Arg His Glu Gln 4175 4180 4185 Arg Ser Ser Arg Gly Gln His Gly Ser Gly Tyr Tyr Tyr Glu Gln 4190 4195 4200 Glu His Ser Glu Glu Glu Ser Asp Ser Gln His Gln Gln Gly His 4205 4210 4215 Gln His Glu Gln Gln Arg Gly His Gln His Gln His Gln His Gln 4220 4225 4230 His Glu His Glu Gln Pro Glu Ser Gly His Arg Gln Gln Gln Ser 4235 4240 4245 Ser Gly Arg Gly His Gln Gly Ala His Gln Glu Gln Gly Arg Asp 4250 4255 4260 Ser Ala Arg Ser Arg Gly Ser Asn Gln Gly His Ser Ser Ser Arg 4265 4270 4275 His Gln Ala Asp Ser Pro Arg Val Ser Ala Arg Ser Gly Ser Gly 4280 4285 4290 Gly Arg Gly Gln Ser Pro Asp Gly Ser Gly Arg Ser Ser Asn Arg 4295 4300 4305 Arg Asp Arg Pro Arg Gln Pro Ser Ala Ser Gln Ser Ser Asp Ser 4310 4315 4320 Gln Val His Ser Gly Val Gln Val Glu Ala Gln Arg Gly Gln Ser 4325 4330 4335 Ser Ser Ala Asn Arg Arg Ala Gly Ser Ser Ser Gly Ser Gly Val 4340 4345 4350 Gln Ser Ala Ala Ala Ser Gly Gln Gly Gly Tyr Glu Ser Ile Phe 4355 4360 4365 Thr Ala Lys His Leu Asp Phe Asn Gln Ser His Ser Tyr Tyr Tyr 4370 4375 4380 Tyr <210> 13 <211> 554 <212> PRT <213> Mus musculus <400> 13 Met Ser Ala Leu Leu Glu Ser Ile Thr Ser Met Ile Glu Ile Phe Gln 1 5 10 15 Gln Tyr Ser Thr Ser Asp Lys Glu Glu Glu Thr Leu Ser Lys Glu Glu 20 25 30 Leu Lys Glu Leu Leu Glu Gly Gln Leu Gln Ala Val Leu Lys Asn Pro 35 40 45 Asp Asp Gln Asp Ile Ala Glu Val Phe Met Gln Met Leu Asp Val Asp 50 55 60 His Asp Asp Lys Leu Asp Phe Ala Glu Tyr Leu Leu Leu Val Leu Lys 65 70 75 80 Leu Ala Lys Ala Tyr Tyr Glu Ala Ser Lys Asn Glu Ser Phe Gln Thr 85 90 95 His Gly Ser Asn Gly Arg Ser Lys Thr Asp Tyr Lys Gly Leu Glu Glu 100 105 110 Glu Gly Glu Glu Gly Asn Glu Gln Asn Leu Arg Arg Arg His Gly Gly 115 120 125 Thr Asp Gly Lys Arg Lys Ser Asp Arg Thr Arg Ser Pro Asn Gly Lys 130 135 140 Arg Gly Lys Arg Gln Glu Ser Arg Cys Arg Ser Glu Gly Lys Asp Lys 145 150 155 160 His Arg Arg Glu Pro Glu Lys His Arg His Gln Gln Asp Ser Lys Arg 165 170 175 Lys Gln Arg His Gly Ser Gly Ser Thr Glu Arg Lys Asp Asn Arg Asn 180 185 190 Lys Lys Asn Arg Gln Ser Lys Glu Arg Asn Tyr Asp Glu Ile Tyr Asp 195 200 205 Asn Gly Lys Tyr Asn Glu Asp Trp Glu Ala Ser Tyr Asn Asn Cys Tyr 210 215 220 Tyr Lys Thr Gln Asn Thr Thr Leu Asp Gln Arg Glu Gly Asn Arg Arg 225 230 235 240 Pro Arg Ala Asp Ser Gln Lys Glu Pro Gln Ser Phe His Gly Gln Ala 245 250 255 Asp Asn Ser Asp Ser Glu Gly Gly Arg Gln Gln Ser His Ser Lys Pro 260 265 270 Ser Pro Val Arg Ala Asp Gln Arg Arg Ser Arg Ala Gly Gln Ala Gly 275 280 285 Ser Ser Lys Val Ser Ala Arg Ser Gly Ser Gly Gly Arg Gly Gln Ser 290 295 300 Pro Asp Gly Ser Gly Arg Ser Ser Asn Arg Arg Asp Arg Pro Arg Gln 305 310 315 320 Pro Ser Pro Ser Gln Ser Ser Asp Ser Gln Val His Ser Gly Val Gln 325 330 335 Val Glu Gly Arg Arg Gly Gln Ser Ser Ser Ala Asn Arg Arg Ala Gly 340 345 350 Ser Ser Ser Gly Ser Gly Val Gln Gly Ala Ser Ala Gly Gly Leu Ala 355 360 365 Ala Asp Ala Ser Arg Arg Thr Gly Ala Leu Gln Gly Gln Ala Ser Ala 370 375 380 Gln Gly Arg Ala Gly Ser Gln Gly Gln Ala Gln Gly Arg Val Gly Ser 385 390 395 400 Ser Ala Asp Arg Gln Gly Arg Arg Gly Val Ser Glu Ser Gln Ala Ser 405 410 415 Asp Ser Glu Gly Gln Ser Asp Phe Ser Glu Gly Gln Ala Val Gly Ala 420 425 430 His Arg Gln Ser Gly Ala Gly Gln Arg His Glu Gln Arg Ser Ser Arg 435 440 445 Gly Gln Tyr Gly Ser Gly Phe Tyr Pro Val Tyr Tyr Tyr Tyr Glu Gln 450 455 460 Glu His Ser Glu Glu Glu Ser Asp Ser Gln His Gly His Gln His Glu 465 470 475 480 Gln Gln Arg Gly His Gln His Gln His Gln His Glu His Glu Gln Pro 485 490 495 Glu Ser Gly His Arg Gln Gln Gln Ser Ser Gly Arg Gly His Gln Gly 500 505 510 Ala His Gln Glu Gln Gly Arg Asp Ser Ala Arg Ser Arg Gly Ser Asn 515 520 525 Gln Gly His Ser Ser Ser Arg His Gln Ala Asp Ser Pro Arg Val Ser 530 535 540 Val Arg Ser Gly Ser Gly Gly Arg Gly Gln 545 550 <210> 14 <211> 336 <212> PRT <213> Mus musculus <400> 14 Pro Asp Gly Ser Gly Arg Ser Ser Asn Arg Arg Asp Arg Pro Arg Gln 1 5 10 15 Leu Ser Pro Ser Gln Ser Ser Asp Ser Gln Val His Ser Gly Val Gln 20 25 30 Val Glu Gly Arg Arg Gly His Ser Ser Ser Ala Asn Arg Arg Ala Gly 35 40 45 Ser Ser Ser Gly Ser Gly Val Gln Gly Ala Ser Ala Gly Gly Leu Ala 50 55 60 Ala Asp Ala Ser Arg Arg Ser Gly Ala Arg Gln Gly Gln Ala Ser Ala 65 70 75 80 Gln Gly Arg Ala Gly Ser Gln Gly Gln Ala Gln Gly Arg Val Ser Ser 85 90 95 Ser Ala Asp Arg Gln Gly Arg Arg Gly Val Ser Glu Ser Arg Ala Ser 100 105 110 Asp Ser Glu Gly His Ser Asp Phe Ser Glu Gly Gln Ala Val Gly Ala 115 120 125 His Arg Gln Ser Gly Ala Gly Gln Arg His Glu Gln Arg Ser Ser Arg 130 135 140 Gly Gln His Gly Ser Gly Tyr Tyr Tyr Glu Gln Glu His Ser Glu Glu 145 150 155 160 Glu Ser Asp Ser Gln His Gln His Gly His Gln His Glu Gln Gln Arg 165 170 175 Gly His Gln His Gln His Gln His Gln His Glu His Glu Gln Pro Glu 180 185 190 Ser Gly His Arg Gln Gln Gln Ser Ser Gly Arg Gly His Gln Gly Ala 195 200 205 His Gln Glu Gln Gly Arg Asp Ser Ala Arg Pro Arg Gly Ser Asn Gln 210 215 220 Gly His Ser Ser Ser Arg His Gln Ala Asp Ser Pro Arg Val Ser Ala 225 230 235 240 Arg Ser Gly Ser Gly Gly Arg Gly Gln Ser Pro Asp Ala Ser Gly Arg 245 250 255 Ser Ser Asn Arg Arg Asp Arg Pro Arg Gln Pro Ser Pro Ser Gln Ser 260 265 270 Ser Asp Ser Gln Val His Ser Gly Val Gln Val Glu Ala Gln Arg Gly 275 280 285 Gln Ser Ser Ser Ala Asn Arg Arg Ala Gly Ser Ser Ser Gly Ser Gly 290 295 300 Val Gln Gly Ala Ala Ala Ser Gly Gln Gly Gly Tyr Glu Ser Ile Phe 305 310 315 320 Thr Ala Lys His Leu Asp Phe Asn Gln Ser His Ser Tyr Tyr Tyr Tyr 325 330 335 <210> 15 <211> 250 <212> PRT <213> Mus musculus <400> 15 Gly Gly Leu Ala Ala Asp Ala Ser Arg Arg Ser Gly Ala Leu Gln Gly 1 5 10 15 Gln Ala Ser Ala Gln Gly Arg Ala Gly Ser Gln Gly Gln Ala Gln Gly 20 25 30 Arg Val Gly Ser Ser Ala Asp Arg Gln Gly Arg Arg Gly Val Ser Glu 35 40 45 Ser Gln Ala Ser Asp Ser Glu Gly His Ser Asp Phe Ser Glu Gly Gln 50 55 60 Ala Val Gly Ala His Arg Gln Ser Gly Ala Gly Gln Arg His Glu Gln 65 70 75 80 Arg Ser Ser Arg Gly Gln His Gly Ser Gly Tyr Tyr Tyr Glu Gln Glu 85 90 95 His Ser Glu Glu Glu Ser Asp Ser Gln His Gln His Gly His Gln His 100 105 110 Glu Gln Gln Arg Gly His Gln His Gln His Gln His Gln His Glu His 115 120 125 Glu Gln Pro Glu Ser Gly His Arg Gln Gln Gln Ser Ser Gly Arg Gly 130 135 140 His Gln Gly Ala His Gln Glu Gln Gly Arg Asp Ser Ala Arg Ser Arg 145 150 155 160 Gly Ser Asn Gln Gly His Ser Ser Ser Arg His Gln Ala Asp Ser Pro 165 170 175 Arg Val Ser Ala Arg Ser Gly Ser Gly Gly Arg Gly Gln Ser Pro Asp 180 185 190 Ala Ser Gly Arg Ser Ser Asn Arg Arg Asp Arg Pro Arg Gln Pro Ser 195 200 205 Pro Ser Gln Ser Ser Asp Ser Gln Val His Ser Gly Val Gln Val Glu 210 215 220 Gly Arg Arg Gly Gln Ser Ser Ser Ala Asn Arg Arg Ala Gly Ser Ser 225 230 235 240 Ser Ser Ser Gly Val Gln Gly Ala Ser Ala 245 250 <210> 16 <211> 2642 <212> PRT <213> Mus musculus <400> 16 Met Ser Thr Asn Ser Pro Ser Pro Ala Thr Gly Ser Ser Ser Ser Ala 1 5 10 15 Asp Ser Pro Arg Val Ser Ala Arg Ser Gly Ser Gly Gly Arg Gly Gln 20 25 30 Ser Pro Asp Ala Ser Gly Arg Ser Ser Asn Arg Arg Asp Arg Pro Arg 35 40 45 Gln Pro Ser Pro Ser Gln Ser Ser Asp Ser Gln Val His Ser Gly Val 50 55 60 Gln Val Glu Gly Arg Arg Gly Gln Ser Ser Ser Ala Asn Arg Arg Ala 65 70 75 80 Gly Ser Ser Ser Gly Ser Gly Val Gln Gly Ala Ser Ala Gly Gly Leu 85 90 95 Ala Ala Asp Ala Ser Arg Arg Ser Gly Ala Arg Gln Gly Gln Ala Ser 100 105 110 Ala Gln Gly Arg Ala Gly Ser Gln Gly Gln Ala Gln Gly Arg Val Gly 115 120 125 Ser Ser Ala Asp Arg Gln Gly Arg Arg Gly Val Ser Glu Ser Gln Ala 130 135 140 Ser Asp Ser Glu Gly His Ser Asp Phe Ser Glu Gly Gln Ala Val Gly 145 150 155 160 Ala His Arg Gln Ser Gly Ala Gly Gln Arg His Glu Gln Arg Ser Ser 165 170 175 Arg Gly Gln His Gly Ser Gly Phe Tyr Pro Val Tyr Tyr Tyr Tyr Glu 180 185 190 Gln Glu His Ser Glu Glu Glu Ser Asp Ser Gln His Gln His Gly His 195 200 205 Gln His Glu Gln Gln Arg Gly His Gln His Gln His Gln His Gln His 210 215 220 Glu His Glu Gln Pro Glu Ser Gly His Arg Gln Gln Gln Ser Ser Gly 225 230 235 240 Arg Gly His Gln Gly Ala His Gln Glu Gln Gly Arg Asp Ser Ala Arg 245 250 255 Ser Arg Gly Ser Asn Gln Gly His Ser Ser Ser Arg His Gln Ala Asp 260 265 270 Ser Pro Arg Val Ser Ala Arg Ser Gly Ser Gly Gly Arg Gly Gln Ser 275 280 285 Pro Asp Ala Ser Gly Arg Ser Ser Asn Arg Arg Asp Arg Pro Arg Gln 290 295 300 Pro Ser Pro Ser Gln Ser Ser Asp Ser Gln Val His Ser Gly Val Gln 305 310 315 320 Val Glu Gly Arg Arg Gly Gln Ser Ser Ser Ala Asn Arg Arg Ala Gly 325 330 335 Ser Ser Ser Gly Ser Gly Val Gln Gly Ala Ser Ala Gly Gly Leu Ala 340 345 350 Ala Asp Ala Ser Arg Arg Ser Gly Ala Arg Gln Gly Gln Ala Ser Ala 355 360 365 Gln Gly Arg Ala Gly Ser Gln Gly Gln Ala Gln Gly Arg Val Gly Ser 370 375 380 Ser Ala Asp Arg Gln Gly Arg Arg Gly Val Ser Glu Ser Gln Ala Ser 385 390 395 400 Asp Ser Glu Gly His Ser Asp Phe Ser Glu Gly Gln Ala Val Gly Ala 405 410 415 His Arg Gln Ser Gly Ala Gly Gln Arg His Glu Gln Arg Ser Ser Arg 420 425 430 Gly Gln His Gly Ser Gly Tyr Tyr Tyr Glu Gln Glu His Ser Glu Glu 435 440 445 Glu Ser Asp Ser Gln His Gln His Ser His Gln His Glu Gln Gln Arg 450 455 460 Gly His Gln His Gln His Gln His Gln His Glu His Glu Gln Pro Glu 465 470 475 480 Ser Gly His Arg Gln Gln Gln Phe Ser Gly Arg Gly His Gln Gly Ala 485 490 495 His Gln Glu Gln Gly Arg Asp Ser Ala Arg Ser Arg Gly Ser Asn Gln 500 505 510 Gly His Ser Ser Ser Arg His Gln Ala Asp Ser Pro Arg Val Ser Ala 515 520 525 Arg Ser Gly Ser Gly Gly Arg Gly Gln Ser Pro Asp Ala Ser Gly Arg 530 535 540 Ser Ser Asn Arg Arg Asp Arg Pro Arg Gln Pro Ser Pro Ser Gln Ser 545 550 555 560 Ser Asp Ser Gln Val His Ser Gly Val Gln Val Glu Gly Arg Arg Gly 565 570 575 Gln Ser Ser Ser Ala Asn Arg Arg Ala Gly Ser Ser Ser Gly Ser Gly 580 585 590 Val Gln Gly Ala Ser Ala Gly Gly Leu Ala Ala Asp Ala Ser Arg Arg 595 600 605 Ser Gly Ala Arg Gln Gly Gln Ala Ser Ala Gln Gly Arg Ala Gly Ser 610 615 620 Gln Gly Gln Ala Gln Gly Arg Val Gly Ser Ser Ala Asp Arg Gln Gly 625 630 635 640 Arg Arg Gly Val Ser Glu Ser Gln Ala Ser Asp Ser Glu Gly His Ser 645 650 655 Asp Phe Ser Glu Gly Gln Ala Val Gly Ala His Arg Gln Ser Gly Ala 660 665 670 Gly Gln Arg His Glu Gln Arg Ser Ser Arg Gly Gln His Gly Ser Gly 675 680 685 Phe Tyr Pro Val Tyr Tyr Tyr Tyr Glu Gln Glu His Ser Glu Glu Glu 690 695 700 Ser Asp Ser Gln His Gln His Gly His Gln His Glu Gln Gln Arg Gly 705 710 715 720 His Gln His Gln His Gln His Gln His Glu His Glu Gln Pro Glu Ser 725 730 735 Gly His Arg Gln Gln Gln Ser Ser Gly Arg Gly His Gln Gly Ala His 740 745 750 Gln Glu Gln Gly Arg Asp Ser Ala Arg Ser Arg Gly Ser Asn Gln Gly 755 760 765 His Ser Ser Ser Arg His Gln Ala Asp Ser Pro Arg Val Ser Ala Arg 770 775 780 Ser Gly Ser Gly Gly Arg Gly Gln Ser Pro Asp Ala Ser Gly Arg Ser 785 790 795 800 Ser Asn Arg Arg Asp Arg Pro Arg Gln Pro Ser Pro Ser Gln Ser Ser 805 810 815 Asp Ser Gln Val His Ser Gly Val Gln Val Glu Gly Arg Arg Gly Gln 820 825 830 Ser Ser Ser Ala Asn Arg Arg Ala Gly Ser Ser Ser Gly Ser Gly Val 835 840 845 Gln Gly Ala Ser Ala Gly Gly Leu Ala Ala Asp Ala Ser Arg Arg Ser 850 855 860 Gly Ala Arg Gln Gly Gln Ala Ser Ala Gln Gly Arg Ala Gly Ser Gln 865 870 875 880 Gly Gln Ala Gln Gly Arg Val Gly Ser Ser Ala Asp Arg Gln Gly Arg 885 890 895 Arg Gly Val Ser Glu Ser Gln Ala Ser Asp Ser Glu Gly His Ser Asp 900 905 910 Phe Ser Glu Gly Gln Ala Val Gly Ala His Arg Gln Ser Gly Ala Gly 915 920 925 Gln Arg His Glu Gln Arg Ser Ser Arg Ser Gln His Gly Ser Gly Tyr 930 935 940 Tyr Tyr Glu Gln Glu His Ser Glu Glu Glu Ser Asp Ser Gln His Gln 945 950 955 960 His Ser His Gln His Glu Gln Gln Arg Gly His Gln His Gln His Gln 965 970 975 His Gln His Glu His Glu Gln Pro Glu Ser Gly His Arg Gln Gln Gln 980 985 990 Ser Ser Gly Arg Gly Asn Gln Gly Ala His Gln Glu Gln Gly Arg Asp 995 1000 1005 Ser Ala Arg Ser Arg Gly Ser Asn Gln Gly His Ser Ser Ser Arg 1010 1015 1020 His Gln Ala Asp Ser Pro Arg Val Ser Ala Arg Ser Gly Ser Gly 1025 1030 1035 Gly Arg Gly Gln Ser Pro Asp Ala Ser Gly Arg Ser Ser Asn Arg 1040 1045 1050 Arg Asp Arg Pro Arg Gln Pro Ser Pro Ser Gln Ser Ser Asp Ser 1055 1060 1065 His Val His Ser Gly Val Gln Val Glu Gly Arg Arg Gly Gln Ser 1070 1075 1080 Ser Ser Ala Asn Arg Arg Ala Gly Ser Ser Ser Gly Ser Gly Val 1085 1090 1095 Gln Gly Ala Ser Ala Gly Gly Leu Ala Ala Asp Ala Ser Arg Arg 1100 1105 1110 Ser Gly Ala Arg Gln Gly Gln Ala Ser Ala Gln Gly Arg Ala Gly 1115 1120 1125 Ser Gln Gly Gln Ala Gln Gly Arg Val Gly Ser Ser Ala Asp Arg 1130 1135 1140 Gln Gly Arg Arg Gly Val Ser Glu Ser Gln Ala Ser Asp Ser Glu 1145 1150 1155 Gly His Ser Asp Phe Ser Glu Gly Gln Ala Val Gly Ala His Arg 1160 1165 1170 Gln Ser Gly Ala Gly Gln Arg His Glu Gln Arg Ser Ser Arg Gly 1175 1180 1185 Gln His Gly Ser Gly Phe Tyr Pro Val Tyr Tyr Tyr Tyr Glu Gln 1190 1195 1200 Glu His Ser Glu Glu Glu Ser Asp Ser Gln His Gln His Gly His 1205 1210 1215 Gln His Glu Gln Gln Arg Gly His Gln His Gln His Gln His Gln 1220 1225 1230 His Glu His Glu Gln Pro Glu Ser Gly His Arg Gln Gln Gln Ser 1235 1240 1245 Ser Gly Arg Gly His Gln Gly Ala His Gln Glu Gln Gly Arg Asp 1250 1255 1260 Ser Ala Arg Ser Arg Gly Ser Asn Gln Gly His Ser Ser Ser Arg 1265 1270 1275 His Gln Ala Asp Ser Pro Arg Val Ser Ala Arg Ser Gly Ser Gly 1280 1285 1290 Gly Arg Gly Gln Ser Pro Asp Ala Ser Gly Arg Ser Ser Asn Arg 1295 1300 1305 Arg Asp Arg Ser Arg Gln Pro Ser Pro Ser Gln Ser Ser Asp Ser 1310 1315 1320 Gln Val His Ser Gly Val Gln Val Glu Gly Arg Arg Gly Gln Ser 1325 1330 1335 Ser Ser Ala Asn Arg Arg Ala Gly Ser Ser Ser Gly Ser Gly Val 1340 1345 1350 Gln Gly Ala Ser Ala Gly Gly Leu Ala Ala Asp Ala Ser Arg Arg 1355 1360 1365 Ser Gly Ala Arg Gln Gly Gln Ala Ser Ala Gln Gly Arg Ala Gly 1370 1375 1380 Ser Gln Gly Gln Ala Gln Gly Arg Val Gly Ser Ser Ala Asp Arg 1385 1390 1395 Gln Gly Arg Arg Gly Val Ser Glu Ser Gln Ala Ser Asp Ser Glu 1400 1405 1410 Gly His Ser Asp Phe Ser Glu Gly Gln Ala Val Gly Ala His Arg 1415 1420 1425 Gln Ser Gly Ala Gly Gln Arg His Glu Gln Arg Ser Ser Arg Gly 1430 1435 1440 Gln His Gly Ser Gly Phe Tyr Pro Val Tyr Tyr Tyr Tyr Glu Gln 1445 1450 1455 Glu His Ser Glu Glu Glu Ser Asp Ser Gln His Gln His Gly His 1460 1465 1470 Gln His Glu Gln Gln Arg Gly His Gln His Gln His Gln His Glu 1475 1480 1485 His Glu Gln Pro Glu Ser Gly His Arg Gln Gln Gln Ser Ser Gly 1490 1495 1500 Arg Gly His Gln Gly Ala His Gln Glu Gln Gly Arg Asp Ser Ala 1505 1510 1515 Arg Ser Arg Gly Ser Asn Gln Gly His Ser Ser Ser Arg His Gln 1520 1525 1530 Ala Asp Ser Pro Arg Val Ser Ala Arg Ser Gly Ser Gly Gly Arg 1535 1540 1545 Gly Gln Ser Pro Asp Ala Ser Gly Arg Ser Ser Asn Arg Arg Asp 1550 1555 1560 Arg Pro Arg Gln Pro Ser Pro Ser Gln Ser Ser Asp Ser Gln Val 1565 1570 1575 His Ser Gly Val Gln Val Glu Gly Arg Arg Gly Gln Ser Ser Ser 1580 1585 1590 Ala Asn Arg Arg Ala Gly Ser Ser Ser Gly Ser Gly Val Gln Gly 1595 1600 1605 Ala Ser Ala Gly Gly Leu Ala Ala Asp Ala Ser Arg Arg Ser Gly 1610 1615 1620 Ala Leu Gln Gly Gln Ala Ser Ala Gln Gly Arg Ala Gly Ser Gln 1625 1630 1635 Gly Gln Ala Gln Gly Arg Val Gly Ser Ser Ala Asp Arg Gln Gly 1640 1645 1650 Arg Arg Gly Val Ser Glu Ser Gln Ala Ser Asp Ser Glu Gly His 1655 1660 1665 Ser Asp Phe Ser Glu Gly Gln Ala Val Gly Ala His Arg Gln Ser 1670 1675 1680 Gly Ala Gly Gln Arg His Glu Gln Arg Ser Ser Arg Gly Gln His 1685 1690 1695 Gly Ser Gly Tyr Tyr Tyr Glu Gln Glu His Ser Glu Glu Glu Ser 1700 1705 1710 Asp Ser Gln His Gln His Gly His Gln His Glu Gln Gln Arg Gly 1715 1720 1725 His Gln His Gln His Gln His Gln His Glu His Glu Gln Pro Glu 1730 1735 1740 Ser Gly His Arg Gln Gln Gln Ser Ser Gly Arg Gly His Gln Gly 1745 1750 1755 Ala His Gln Glu Gln Gly Arg Asp Ser Ala Arg Ser Arg Gly Ser 1760 1765 1770 Asn Gln Gly His Ser Ser Ser Arg His Gln Ala Asp Ser Pro Arg 1775 1780 1785 Val Ser Ala Arg Ser Gly Ser Gly Gly Arg Gly Gln Ser Pro Asp 1790 1795 1800 Ala Ser Gly Arg Ser Ser Asn Arg Arg Asp Arg Pro Arg Gln Pro 1805 1810 1815 Ser Pro Ser Gln Ser Ser Asp Ser Gln Val His Ser Gly Val Gln 1820 1825 1830 Val Glu Gly Arg Arg Gly Gln Ser Ser Ser Ala Asn Arg Arg Ala 1835 1840 1845 Gly Ser Ser Ser Gly Ser Gly Val Gln Gly Ala Ser Ala Gly Gly 1850 1855 1860 Leu Ala Ala Asp Ala Ser Arg Arg Ser Gly Ala Arg Gln Gly Gln 1865 1870 1875 Ala Ser Ala Gln Gly Arg Ala Gly Ser Gln Gly Gln Ala Gln Gly 1880 1885 1890 Arg Val Gly Ser Ser Ala Asp Arg Gln Gly Arg Arg Gly Val Ser 1895 1900 1905 Gly Ser Gln Ala Ser Asp Ser Glu Gly His Ser Asp Phe Ser Glu 1910 1915 1920 Gly Gln Ala Val Gly Ala His Arg Gln Ser Gly Ala Gly Gln Arg 1925 1930 1935 His Glu Gln Arg Ser Ser Arg Gly Gln His Gly Ser Gly Tyr Tyr 1940 1945 1950 Tyr Glu Gln Glu His Ser Glu Glu Glu Ser Asp Ser Gln His Gln 1955 1960 1965 His Gly His Gln His Glu Gln Gln Arg Gly His Gln His Gln His 1970 1975 1980 Gln His Gln His Glu His Glu Gln Pro Glu Ser Gly His Arg Gln 1985 1990 1995 Gln Gln Phe Ser Gly Arg Gly His Gln Gly Ala His Gln Glu Gln 2000 2005 2010 Gly Arg Asp Ser Ala Arg Ser Arg Gly Ser Asn Gln Gly His Ser 2015 2020 2025 Ser Ser Arg His Gln Ala Asp Ser Pro Arg Val Ser Ala Arg Ser 2030 2035 2040 Gly Ser Gly Gly Arg Gly Gln Ser Pro Asp Ala Ser Gly Arg Ser 2045 2050 2055 Ser Asn Arg Arg Asp Arg Pro Arg Gln Pro Ser Pro Ser Gln Ser 2060 2065 2070 Ser Asp Ser Gln Val His Ser Gly Val Gln Val Glu Gly Arg Arg 2075 2080 2085 Gly Gln Ser Ser Ser Ala Asn Arg Arg Ala Gly Ser Ser Ser Ser 2090 2095 2100 Ser Gly Val Gln Gly Ala Ser Ala Gly Gly Leu Ala Ala Asp Ala 2105 2110 2115 Ser Arg Arg Ser Gly Ala Arg Gln Gly Gln Ala Ser Ala Gln Gly 2120 2125 2130 Arg Ala Gly Ser Gln Gly Gln Ala Gln Gly Arg Val Gly Ser Ser 2135 2140 2145 Ala Asp Arg Gln Gly Arg Arg Gly Val Ser Glu Ser Gln Ala Ser 2150 2155 2160 Asp Ser Glu Gly His Ser Asp Phe Ser Glu Gly Gln Ala Val Gly 2165 2170 2175 Ala His Arg Gln Ser Gly Ala Gly Gln Arg His Glu Gln Arg Ser 2180 2185 2190 Ser Arg Gly Gln His Gly Ser Gly Phe Tyr Pro Val Tyr Tyr Tyr 2195 2200 2205 Tyr Glu Gln Glu His Ser Glu Glu Glu Ser Asp Ser Gln His Gln 2210 2215 2220 His Gly His Gln His Glu Gln Gln Arg Gly His Gln His Gln His 2225 2230 2235 Gln His Gln His Glu His Glu Gln Pro Glu Ser Gly His Arg Gln 2240 2245 2250 Gln Gln Phe Ser Gly Arg Gly His Gln Gly Ala His Gln Glu Gln 2255 2260 2265 Gly Arg Asp Ser Ala Arg Ser Arg Gly Ser Asn Gln Gly His Ser 2270 2275 2280 Ser Ser Arg His Gln Ala Asp Ser Pro Arg Val Ser Ala Arg Ser 2285 2290 2295 Gly Ser Gly Gly Arg Gly Gln Ser Pro Asp Ala Ser Gly Arg Ser 2300 2305 2310 Ser Asn Arg Arg Asp Arg Pro Arg Gln Pro Ser Pro Ser Gln Ser 2315 2320 2325 Ser Asp Ser Gln Val His Ser Gly Val Gln Val Glu Gly Arg Arg 2330 2335 2340 Gly Gln Ser Ser Ser Ala Asn Arg Arg Ala Gly Ser Ser Ser Gly 2345 2350 2355 Ser Gly Val Gln Gly Ala Ser Ala Gly Gly Leu Ala Ala Asp Ala 2360 2365 2370 Ser Arg Arg Ser Gly Ala Arg Gln Gly Gln Ala Ser Ala Gln Gly 2375 2380 2385 Arg Ala Gly Ser Gln Gly Gln Ala Gln Gly Arg Val Gly Ser Ser 2390 2395 2400 Ala Asp Arg Gln Gly Arg Arg Gly Val Ser Glu Ser Gln Ala Ser 2405 2410 2415 Asp Ser Glu Gly His Ser Asp Phe Ser Glu Gly Gln Ala Val Gly 2420 2425 2430 Ala His Arg Gln Ser Gly Ala Gly Gln Arg His Glu Gln Arg Ser 2435 2440 2445 Ser Arg Gly Gln His Gly Ser Gly Tyr Tyr Tyr Glu Gln Glu His 2450 2455 2460 Ser Glu Glu Glu Ser Asp Ser Gln His Gln Gln Gly His Gln His 2465 2470 2475 Glu Gln Gln Arg Gly His Gln His Gln His Gln His Gln His Glu 2480 2485 2490 His Glu Gln Pro Glu Ser Gly His Arg Gln Gln Gln Ser Ser Gly 2495 2500 2505 Arg Gly His Gln Gly Ala His Gln Glu Gln Gly Arg Asp Ser Ala 2510 2515 2520 Arg Ser Arg Gly Ser Asn Gln Gly His Ser Ser Ser Arg His Gln 2525 2530 2535 Ala Asp Ser Pro Arg Val Ser Ala Arg Ser Gly Ser Gly Gly Arg 2540 2545 2550 Gly Gln Ser Pro Asp Gly Ser Gly Arg Ser Ser Asn Arg Arg Asp 2555 2560 2565 Arg Pro Arg Gln Pro Ser Ala Ser Gln Ser Ser Asp Ser Gln Val 2570 2575 2580 His Ser Gly Val Gln Val Glu Ala Gln Arg Gly Gln Ser Ser Ser 2585 2590 2595 Ala Asn Arg Arg Ala Gly Ser Ser Ser Gly Ser Gly Val Gln Ser 2600 2605 2610 Ala Ala Ala Ser Gly Gln Gly Gly Tyr Glu Ser Ile Phe Thr Ala 2615 2620 2625 Lys His Leu Asp Phe Asn Gln Ser His Ser Tyr Tyr Tyr Tyr 2630 2635 2640 <210> 17 <211> 336 <212> PRT <213> Mus musculus <400> 17 Pro Asp Gly Ser Gly Arg Ser Ser Asn Arg Arg Asp Arg Pro Arg Gln 1 5 10 15 Leu Ser Pro Ser Gln Ser Ser Asp Ser Gln Val His Ser Gly Val Gln 20 25 30 Val Glu Gly Arg Arg Gly His Ser Ser Ser Ala Asn Arg Arg Ala Gly 35 40 45 Ser Ser Ser Gly Ser Gly Val Gln Gly Ala Ser Ala Gly Gly Leu Ala 50 55 60 Ala Asp Ala Ser Arg Arg Ser Gly Ala Arg Gln Gly Gln Ala Ser Ala 65 70 75 80 Gln Gly Arg Ala Gly Ser Gln Gly Gln Ala Gln Gly Arg Val Ser Ser 85 90 95 Ser Ala Asp Arg Gln Gly Arg Arg Gly Val Ser Glu Ser Arg Ala Ser 100 105 110 Asp Ser Glu Gly His Ser Asp Phe Ser Glu Gly Gln Ala Val Gly Ala 115 120 125 His Arg Gln Ser Gly Ala Gly Gln Arg His Glu Gln Arg Ser Ser Arg 130 135 140 Gly Gln His Gly Ser Gly Tyr Tyr Tyr Glu Gln Glu His Ser Glu Glu 145 150 155 160 Glu Ser Asp Ser Gln His Gln His Gly His Gln His Glu Gln Gln Arg 165 170 175 Gly His Gln His Gln His Gln His Gln His Glu His Glu Gln Pro Glu 180 185 190 Ser Gly His Arg Gln Gln Gln Ser Ser Gly Arg Gly His Gln Gly Ala 195 200 205 His Gln Glu Gln Gly Arg Asp Ser Ala Arg Pro Arg Gly Ser Asn Gln 210 215 220 Gly His Ser Ser Ser Arg His Gln Ala Asp Ser Pro Arg Val Ser Ala 225 230 235 240 Arg Ser Gly Ser Gly Gly Arg Gly Gln Ser Pro Asp Ala Ser Gly Arg 245 250 255 Ser Ser Asn Arg Arg Asp Arg Pro Arg Gln Pro Ser Pro Ser Gln Ser 260 265 270 Ser Asp Ser Gln Val His Ser Gly Val Gln Val Glu Ala Gln Arg Gly 275 280 285 Gln Ser Ser Ser Ala Asn Arg Arg Ala Gly Ser Ser Ser Gly Ser Gly 290 295 300 Val Gln Gly Ala Ala Ala Ser Gly Gln Gly Gly Tyr Glu Ser Ile Phe 305 310 315 320 Thr Ala Lys His Leu Asp Phe Asn Gln Ser His Ser Tyr Tyr Tyr Tyr 325 330 335 <210> 18 <211> 357 <212> PRT <213> Mus musculus <400> 18 Met Ser Ala Leu Leu Glu Ser Ile Thr Ser Met Ile Glu Ile Phe Gln 1 5 10 15 Gln Tyr Ser Thr Ser Asp Lys Glu Glu Glu Thr Leu Ser Lys Glu Glu 20 25 30 Leu Lys Glu Leu Leu Glu Gly Gln Leu Gln Ala Val Leu Lys Asn Pro 35 40 45 Asp Asp Gln Asp Ile Ala Glu Val Phe Met Gln Met Leu Asp Val Asp 50 55 60 His Asp Asp Lys Leu Asp Phe Ala Glu Tyr Leu Leu Leu Val Leu Lys 65 70 75 80 Leu Ala Lys Ala Tyr Tyr Glu Ala Ser Lys Asn Glu Ser Phe Gln Thr 85 90 95 His Gly Ser Asn Gly Arg Ser Lys Thr Asp Tyr Lys Gly Leu Glu Glu 100 105 110 Glu Gly Glu Glu Gly Asn Lys Gln Asn Leu Arg Arg Arg His Gly Gly 115 120 125 Thr Asp Gly Lys Arg Lys Ser Asp Arg Thr Arg Ser Pro Asn Gly Lys 130 135 140 Arg Gly Lys Arg Gln Glu Ser Arg Cys Arg Ser Glu Gly Lys Asp Lys 145 150 155 160 His Arg Arg Glu Pro Glu Lys His Arg His Gln Gln Asp Ser Lys Arg 165 170 175 Lys Gln Arg His Gly Ser Gly Ser Thr Glu Arg Lys Asp Asn Arg Asn 180 185 190 Lys Lys Asn Arg Gln Ser Lys Glu Arg Asn Tyr Asp Glu Ile Tyr Asp 195 200 205 Asn Gly Lys Tyr Asn Glu Asp Trp Glu Ala Ser Tyr Asn Asn Cys Tyr 210 215 220 Tyr Lys Thr Gln Asn Thr Thr Leu Asp Gln Arg Glu Gly Asn Arg Arg 225 230 235 240 Pro Arg Ala Asp Ser Gln Lys Glu Pro Gln Ser Ser His Gly Gln Ala 245 250 255 Asp Asn Ser Asp Ser Glu Gly Gly Arg Gln Gln Ser His Ser Lys Pro 260 265 270 Ser Pro Val Arg Ala Asp Gln Arg Arg Ser Arg Ala Gly Gln Ala Gly 275 280 285 Ser Ser Lys Val Ser Ala Arg Ser Gly Ser Gly Gly Arg Gly Gln Ser 290 295 300 Pro Asp Gly Ser Gly Arg Ser Ser Asn Arg Arg Asp Arg Pro Arg Gln 305 310 315 320 Pro Ser Pro Ser Gln Ser Ser Asp Ser Gln Val His Ser Gly Val Gln 325 330 335 Val Glu Gly Arg Arg Gly Gln Ser Ser Ser Ala Asn Arg Arg Ala Gly 340 345 350 Ser Ser Ser Gly Ser 355 <210> 19 <211> 366 <212> domestic worker <213> Rattus norvegicus <400> 19 Met Ser Thr Leu Leu Glu Ser Ile Thr Ser Met Ile Asp Ile Phe Gln 1 5 10 15 Gln Tyr Ser Asn Asn Asp Lys Glu Glu Glu Thr Leu Ser Lys Glu Glu 20 25 30 Leu Lys Glu Leu Leu Glu Gly Glu Leu Gln Ala Val Leu Lys Asn Pro 35 40 45 Asn Asp Gln Asp Ile Ala Glu Val Phe Met Gln Met Leu Asp Val Asp 50 55 60 His Asp Asp Lys Ile Asp Phe Thr Glu Tyr Leu Leu Met Val Leu Lys 65 70 75 80 Leu Ala Gln Ala Tyr Tyr Glu Thr Ser Lys Lys Arg Arg Ser Gln Thr 85 90 95 Lys Glu Ser Gly Lys Arg Asn Glu His Asp Tyr Lys Gly Tyr Glu Glu 100 105 110 Arg Arg Glu Lys Val Gln Arg Arg His Arg Arg Arg Asn Ser Gly Thr 115 120 125 Asp Gly Lys Gln Glu Asn Glu Arg Ser Lys Ser Pro Arg Gly Arg Gly 130 135 140 Asn Lys Arg Arg Gly Ser Ser Thr Ile Ser Glu Glu Ser Asp Thr Asn 145 150 155 160 Arg Asn Ser Asp Thr Glu Asn Lys Arg His His His Gly Ser Asn Arg 165 170 175 Arg Gln Arg Arg Gly Ser Asn Ser Ser Asp Arg Lys Glu Thr Arg Ser 180 185 190 Lys Lys His Arg Glu Val Lys Glu Arg Asn Ala Gly Ile Tyr Asn Asp 195 200 205 Gly Lys Asp Gly Gln Asp Trp Glu Val Asn Tyr Glu Asn Cys Tyr Tyr 210 215 220 Lys Thr Glu Glu Ser Asn Arg Glu Gln Arg Glu Gly Arg Asn His Lys 225 230 235 240 Thr Lys Glu Ser His Ser Glu Ser Glu Ala Ser Gly Gly Gln Ala Gly 245 250 255 Arg Arg Gly Thr Ala Ala Thr Arg His Thr Ser Arg Pro Glu Gln Ser 260 265 270 Pro Asp Thr Ala Gly Arg Thr Gly Ser Ser Arg Gly Gln Gln Ser Ala 275 280 285 Gln Arg His Ala Asp Ser Thr Pro Gly Ser Thr Arg Thr Gly Ser Arg 290 295 300 Gly Arg Gly Glu Ser Pro Ala Gly Gln Gln Ser Pro Asp Arg Ala Arg 305 310 315 320 His Ile Glu Ser Arg Arg Gly Arg Thr Arg Glu Ala Ser Ala Ser Gln 325 330 335 Ser Ser Asp Ser Glu Gly His Ser Gly Ala His Ala Gly Ile Gly Gln 340 345 350 Gly Gln Thr Ser Thr Thr His Arg Arg Ala Gly Ser Ser Ser 355 360 365 <210> 20 <211> 3088 <212> PRT <213> Pan troglodytes <400> 20 Met Ser Thr Leu Leu Glu Asn Ile Phe Ala Ile Ile Asn Leu Phe Lys 1 5 10 15 Gln Tyr Ser Lys Lys Asp Lys Asn Thr Asp Thr Leu Ser Lys Lys Glu 20 25 30 Leu Lys Glu Leu Leu Glu Lys Glu Phe Arg Gln Ile Leu Lys Asn Pro 35 40 45 Asp Asp Pro Asp Met Val Asp Val Phe Met Asp His Leu Asp Ile Asp 50 55 60 His Asn Lys Lys Ile Asp Phe Thr Glu Phe Leu Leu Met Val Phe Lys 65 70 75 80 Leu Ala Gln Ala Tyr Tyr Glu Ser Thr Arg Lys Glu Asn Leu Pro Ile 85 90 95 Ser Gly His Lys His Arg Lys His Ser His His Asp Lys His Glu Asp 100 105 110 Asn Lys Gln Glu Glu Asn Lys Glu Asn Arg Lys Arg Pro Ser Ser Leu 115 120 125 Glu Arg Arg Asn Asn Arg Lys Gly Asn Lys Gly Arg Ser Lys Ser Pro 130 135 140 Arg Glu Thr Gly Gly Lys Arg His Glu Ser Ser Ser Glu Lys Lys Glu 145 150 155 160 Arg Lys Gly Tyr Ser Pro Thr His Arg Glu Glu Glu Tyr Gly Lys Asn 165 170 175 His His Asn Ser Ser Lys Lys Gln Lys Asn Lys Thr Glu Asn Thr Arg 180 185 190 Leu Glu Asp Asn Arg Lys Arg Leu Ser Glu Arg Leu Glu Glu Lys Glu 195 200 205 Asp Asn Glu Glu Gly Gly Tyr Asp Tyr Glu Asn Thr Gly Arg Met Thr 210 215 220 Gln Lys Trp Ile Gln Ser Gly His Ile Ala Thr Tyr Tyr Thr Ile Gln 225 230 235 240 Asp Glu Ala Tyr Asp Thr Thr Asp Asn Leu Leu Glu Glu Asn Lys Ile 245 250 255 Tyr Glu Arg Ser Arg Ser Ser Asp Asp Lys Ser Ser Ser Gln Val Asn 260 265 270 Arg Ser Arg His Glu Asn Thr Ser Gln Val Pro Leu Gln Glu Ser Arg 275 280 285 Thr Arg Lys Arg Arg Gly Ser Arg Val Ser Gln Asp Arg Asp Ser Glu 290 295 300 Gly His Ser Glu Asp Ser Glu Arg His Ser Gly Ser Ala Ser Arg Asn 305 310 315 320 His Pro Gly Ser Ala Arg Gln Gln Ser Arg Asp Gly Ser Arg His Pro 325 330 335 Arg Ser His Asp Glu Asp Arg Ala Ser His Gly His Ser Ala Asp Ser 340 345 350 Ser Arg Gln Ser Gly Thr Arg His Ala Glu Thr Ser Ser Arg Gly Gln 355 360 365 Thr Ala Ser Ser His Glu Gln Ala Arg Ser Ser Pro Gly Glu Arg His 370 375 380 Gly Ser Arg His Gln Gln Ser Ala Asp Ser Ser Arg His Ser Ala Thr 385 390 395 400 Gly Arg Gly Gln Ala Ser Ser Ala Val Ser Asp Arg Gly Pro Arg Gly 405 410 415 Ser Ser Gly Ser Gln Ala Ser Asp Ser Glu Gly His Ser Glu Asn Ser 420 425 430 Asp Thr Gln Ser Val Ser Gly His Gly Lys Ala Gly Pro Arg Gln Gln 435 440 445 Ser His Gln Glu Ser Ala Arg Gly Arg Ser Gly Glu Arg Ser Gly His 450 455 460 Ser Gly Ser Phe Leu Tyr Gln Val Ser Thr His Glu Gln Ser Asp Ser 465 470 475 480 Ala His Gly Arg Thr Gly Thr Ser Thr Gly Gly Arg Gln Gly Ser His 485 490 495 His Glu Gln Ala Arg Asp Ser Ser Arg His Ser Ala Ser Gln Glu Gly 500 505 510 Gln Asp Thr Ile Arg Gly His Pro Gly Ser Ser Arg Gly Gly Arg Gln 515 520 525 Gly Ser His His Glu Gln Leu Val Asn Arg Ser Gly His Ser Gly Ser 530 535 540 His His Ser His Thr Thr Ser Gln Gly Arg Ser Asp Ala Ser His Gly 545 550 555 560 Gln Ser Gly Ser Arg Ser Ala Ser Arg Gln Thr Arg Asn Glu Glu Gln 565 570 575 Ser Gly Asp Gly Ser Arg His Ser Gly Ser Arg His His Glu Ala Ser 580 585 590 Ser Gln Ala Asp Ser Ser Arg His Ser Gln Val Gly Gln Gly Gln Ser 595 600 605 Ser Gly Pro Arg Thr Ser Ser His Gln Gly Ser Ser Val Ser Gln Asp 610 615 620 Ser Asp Ser Glu Gly His Ser Glu Asp Ser Glu Arg Trp Ser Gly Ser 625 630 635 640 Ala Ser Arg Asn His His Gly Ser Ala Arg Glu Gln Ser Arg Asp Gly 645 650 655 Ser Arg His Pro Arg Ser His His Glu Asp Arg Ala Gly His Gly His 660 665 670 Ser Ala Asp Ser Ser Arg Gln Ser Gly Thr Arg His Thr Glu Ser Ser 675 680 685 Ser Arg Gly Gln Ala Val Ser Ser His Glu Gln Ala Arg Ser Ser Pro 690 695 700 Gly Glu Arg His Gly Ser Arg His Gln Gln Ser Ala Asp Ser Ser Arg 705 710 715 720 His Ser Gly Ile Gly His Gly Gln Ala Ser Ser Ala Val Arg Asp Ser 725 730 735 Gly His Arg Gly Ser Ser Gly Ser Gln Ala Ile Asp Ser Glu Gly His 740 745 750 Ser Glu Asp Ser Asp Thr Gln Ser Val Ser Ser His Gly Gln Ala Gly 755 760 765 Pro His Gln Gln Ser His Lys Glu Ser Ala Arg Gly Gln Ser Gly Glu 770 775 780 Ser Ser Gly Arg Ser Gly Ser Phe Leu Tyr Gln Val Ser Ser His Glu 785 790 795 800 Gln Ser Glu Ser Thr Tyr Gly Gln Thr Ala Pro Ser Thr Gly Gly Arg 805 810 815 Gln Gly Ser Arg His Glu Gln Ala Arg Asn Ser Ser Arg His Ser Ala 820 825 830 Ser Gln Asp Gly Gln Asp Thr Ile Arg Gly His Pro Gly Ser Ser Arg 835 840 845 Gly Gly Arg Gln Gly Ser Tyr His Glu Gln Ser Val Asp Arg Ser Gly 850 855 860 His Ser Gly Tyr His His Ser His Thr Thr Pro Gln Gly Arg Ser Asp 865 870 875 880 Ala Ser His Gly Gln Ser Gly Pro Arg Ser Ala Ser Arg Gln Thr Arg 885 890 895 Asn Glu Glu Gln Ser Gly Asp Gly Ser Arg His Ser Gly Ser Arg His 900 905 910 His Glu Pro Ser Thr Arg Ala Ser Ser Ser Arg His Ser Gln Val Gly 915 920 925 Gln Gly Glu Ser Ala Val Ser Lys Thr Ser Arg Arg Gln Gly Ser Ser 930 935 940 Val Ser Gln Asp Arg Asp Ser Glu Gly His Ser Glu Asp Ser Glu Arg 945 950 955 960 Gln Ser Glu Ser Ala Ser Arg Asn His Tyr Gly Ser Ala Arg Glu Gln 965 970 975 Ser Arg His Gly Ser Arg Asn Pro Arg Ser His Gln Glu Asp Arg Ala 980 985 990 Ser His Gly His Ser Ala Glu Ser Ser Arg Gln Ser Gly Thr His His 995 1000 1005 Ala Glu Thr Ser Ser His Gly Gln Ala Ala Ser Ser Gln Glu Gln 1010 1015 1020 Ala Arg Ser Ser Arg Gly Glu Arg His Gly Ser Arg His Gln Gln 1025 1030 1035 Ser Ala Asp Ser Ser Thr Asp Ser Gly Thr Gly Gly Arg Gln Ala 1040 1045 1050 Ser Ser Val Val Gly Asp Ser Gly Asn Arg Gly Ser Ser Gly Ser 1055 1060 1065 Gln Ala Ser Asp Ser Glu Gly His Ser Glu Asp Ser Asp Thr Gln 1070 1075 1080 Ser Val Ser Ala His Gly Gln Ala Gly Pro Arg Gln Gln Ser His 1085 1090 1095 Gln Glu Ser Thr Arg Gly Gln Ser Gly Glu Arg Ser Gly Arg Ser 1100 1105 1110 Gly Ser Phe Leu Tyr Gln Val Ser Thr His Glu Gln Ser Glu Ser 1115 1120 1125 Ala His Gly Arg Thr Gly Pro Ser Thr Gly Gly Arg Gln Arg Ser 1130 1135 1140 Arg His Glu Gln Ala Arg Asp Ser Ser Arg His Ser Ala Ser Gln 1145 1150 1155 Glu Ser Gln Asp Ile Ile His Ala His Pro Gly Ser Ser Arg Gly 1160 1165 1170 Gly Arg Gln Gly Ser His Tyr Glu Gln Ser Val Asp Arg Ser Gly 1175 1180 1185 His Ser Gly Ser His His Ser His Thr Thr Ser Gln Glu Arg Ser 1190 1195 1200 Asn Ala Ser His Gly Gln Ser Gly Ser Arg Ser Ala Ser Arg Gln 1205 1210 1215 Thr Arg Asn Glu Glu Gln Ser Gly Asp Gly Ser Arg His Ser Gly 1220 1225 1230 Ser Arg His His Glu Ala Ser Ser Arg Ala Asp Ser Ser Arg His 1235 1240 1245 Ser Gln Val Gly Gln Gly Gln Ser Ser Gly Pro Arg Thr Ser Arg 1250 1255 1260 Asn Gln Gly Ser Ser Val Ser Gln Asp Ser Asp Ser Gln Gly His 1265 1270 1275 Ser Glu Asp Ser Glu Arg Trp Ser Gly Ser Ala Ser Arg Asn His 1280 1285 1290 His Gly Ser Ala Trp Glu Gln Ser Arg Asp Gly Ser Arg His Pro 1295 1300 1305 Gly Ser His Gln Glu Asp Arg Ala Gly His Gly His Ser Ala Asp 1310 1315 1320 Ser Ser Arg Gln Ser Gly Thr His His Thr Glu Ser Ser Ser Arg 1325 1330 1335 Gly Gln Ala Ala Ser Ser His Glu His Ala Arg Ser Ser Ala Gly 1340 1345 1350 Glu Arg His Gly Ser His His Gln Gln Ser Ala Asp Ser Ser Arg 1355 1360 1365 His Ser Gly Ile Gly His Gly Gln Ala Ser Ser Ala Val Arg Asp 1370 1375 1380 Ser Gly His Arg Gly Ser Ser Gly Ser Gln Ala Ser Asp Ser Glu 1385 1390 1395 Gly His Ser Glu Asp Ser Asp Thr Gln Ser Leu Ser Ala His Gly 1400 1405 1410 Gln Ala Gly Pro His Gln Gln Ser His Gln Glu Ser Thr Arg Gly 1415 1420 1425 Arg Ser Ala Glu Arg Ser Gly Arg Ser Gly Ser Phe Leu Tyr Gln 1430 1435 1440 Val Ser Thr His Glu Gln Ser Glu Ser Ala His Gly Arg Thr Gly 1445 1450 1455 Thr Ser Thr Gly Gly Arg Lys Arg Ser Leu His Glu Gln Ala Arg 1460 1465 1470 Asp Ser Ser Arg His Ser Val Ser Gln Glu Gly Gln Asp Thr Ile 1475 1480 1485 His Gly His Ala Gly Ser Ser Ser Gly Gly Arg Gln Gly Ser His 1490 1495 1500 Tyr Glu Gln Leu Val Asp Arg Ser Gly His Ser Gly Ser His His 1505 1510 1515 Ser His Thr Thr Ser Gln Gly Arg Ser Asp Ala Tyr His Gly Gln 1520 1525 1530 Ser Gly Ser Arg Ser Ala Ser Arg Gln Thr His Asn Asp Glu Gln 1535 1540 1545 Ser Gly Asp Gly Phe Arg His Ser Gly Ser His His His Glu Ala 1550 1555 1560 Ser Ser Arg Ala Asp Ser Ser Arg His Ser Gln Val Gly Gln Gly 1565 1570 1575 Gln Ser Glu Gly Pro Arg Thr Ser Arg His Arg Glu Ser Ser Val 1580 1585 1590 Ser Gln Asp Ser Asp Ser Glu Gly His Ser Glu Asp Ser Glu Arg 1595 1600 1605 Trp Ser Gly Ser Ala Ser Arg Asn His His Gly Ser Ala Arg Glu 1610 1615 1620 Gln Ser Arg Asp Gly Ser Arg His Pro Arg Ser His His Glu Asp 1625 1630 1635 Arg Ala Gly His Gly His Ser Ala Asp Ser Ser Arg Gln Ser Gly 1640 1645 1650 Thr Arg His Thr Gln Thr Ser Ser Gly Gly Gln Ala Ala Ser Ser 1655 1660 1665 His Glu Gln Gly Arg Ser Ser Ala Gly Glu Arg His Gly Ser Arg 1670 1675 1680 His Gln Gln Ser Ala Asp Ser Ser Arg His Ser Gly Ile Gly His 1685 1690 1695 Gly Gln Ala Ser Ser Ala Val Arg Asp Ser Gly His Arg Gly Tyr 1700 1705 1710 Ser Gly Ser Gln Ala Ser Asp Ser Glu Gly His Ser Glu Asp Ser 1715 1720 1725 Asp Thr Gln Ser Val Ser Ala His Gly Gln Ala Gly Ser His Gln 1730 1735 1740 Gln Ser His Gln Glu Ser Ala Arg Gly Arg Ser Gly Glu Arg Ser 1745 1750 1755 Gly Arg Ser Gly Ser Phe Leu Tyr Gln Val Ser Thr His Glu Gln 1760 1765 1770 Ser Glu Ser Ser His Gly Trp Thr Gly Pro Ser Thr Arg Gly Arg 1775 1780 1785 Gln Gly Ser Arg His Glu Gln Ala Gln Asp Ser Ser Arg His Ser 1790 1795 1800 Ala Ser Gln Glu Gly Gln Asp Thr Ile Arg Gly His Pro Gly Pro 1805 1810 1815 Ser Arg Gly Gly Arg Gln Gly Tyr His His Glu Gln Ser Val Asp 1820 1825 1830 Ser Ser Gly His Ser Gly Ser His His Ser His Ile Thr Ser Gln 1835 1840 1845 Gly Ser Ser His Ala Ser His Gly Gln Ser Gly Ser Arg Ser Ala 1850 1855 1860 Ser Arg Thr Thr Arg Asn Asp Glu Gln Ser Val Asp Gly Ser Arg 1865 1870 1875 His Ser Gly Ser Arg His His Glu Ala Ser Thr His Ala Asp Ile 1880 1885 1890 Ser Arg His Ser Gln Ala Gly Gln Gly Gln Ser Glu Gly Ser Arg 1895 1900 1905 Thr Ser Arg Arg Gln Gly Ser Ser Val Ser Gln Asp Ser Asp Ser 1910 1915 1920 Glu Gly His Ser Glu Asp Ser Glu Arg Trp Ser Gly Ser Ala Ser 1925 1930 1935 Arg Asn His Arg Gly Ser Ala Gln Glu Gln Ser Arg Asp Gly Ser 1940 1945 1950 Arg His Pro Arg Ser His His Glu Asp Arg Ala Gly His Gly His 1955 1960 1965 Ser Ala Asp Ser Ser Arg Gln Ser Gly Thr His His Ala Glu Thr 1970 1975 1980 Ser Ser Gly Gly Gln Ala Ala Ser Ser Arg Glu Gln Ala Arg Ser 1985 1990 1995 Ser Pro Gly Glu Arg His Gly Ser Arg His Gln Gln Ser Ala Asp 2000 2005 2010 Ser Ser Arg His Ser Gly Ile Arg Arg Gly Gln Ala Ser Ser Ala 2015 2020 2025 Val Arg Asp Ser Gly His Trp Gly Ser Ser Gly Ser Gln Ala Ser 2030 2035 2040 Asp Ser Glu Gly His Ser Glu Glu Ser Asp Thr Gln Ser Val Ser 2045 2050 2055 Gly His Gly Gln Asp Gly Pro His Gln Gln Ser His Gln Glu Ser 2060 2065 2070 Ala Arg Asp Arg Ser Gly Gly Arg Ser Gly Arg Ser Gly Ser Phe 2075 2080 2085 Leu Tyr Gln Val Ser Thr His Glu Gln Ser Glu Ser Thr His Gly 2090 2095 2100 Gln Thr Gly Thr Ser Thr Gly Gly Arg Gln Gly Ser His His Glu 2105 2110 2115 Gln Ala Arg Asp Ser Ser Arg His Ser Ala Ser Gln Glu Gly Gln 2120 2125 2130 Asp Thr Ile His Ala His Pro Gly Ser Ser Arg Gly Gly Arg Gln 2135 2140 2145 Gly Ser His His Glu Gln Ser Val Asp Thr Ser Gly His Ser Gly 2150 2155 2160 Ser His His Ser His Thr Thr Ser Gln Gly Arg Ser Asp Ala Ser 2165 2170 2175 His Gly Gln Ser Gly Ser Arg Ser Ala Ser Arg Gln Thr Arg Asn 2180 2185 2190 Asp Glu Gln Ser Gly Asp Gly Ser Arg His Ser Gly Ser His His 2195 2200 2205 His Glu Ala Phe Thr Gln Ala Asp Ser Ser Arg His Ser Gln Ser 2210 2215 2220 Gly Gln Gly Glu Ser Ala Gly Ser Arg Arg Ser Arg Arg Gln Gly 2225 2230 2235 Ser Ser Val Ser Gln Asp Ser Asp Ser Gln Gly His Ser Glu Asp 2240 2245 2250 Ser Glu Arg Trp Ser Gly Ser Ala Ser Arg Asn Gln His Gly Ser 2255 2260 2265 Ala Arg Glu Gln Ser Arg Asp Gly Ser Arg His Pro Gly Ser His 2270 2275 2280 Gln Glu Asp Arg Ala Gly His Gly His Ser Ala Asp Ser Ser Arg 2285 2290 2295 Gln Ser Gly Thr Arg His Thr Glu Ser Ser Ser Arg Gly Gln Ala 2300 2305 2310 Ala Ser Ser His Glu Gln Ala Arg Ser Ser Ala Gly Glu Arg His 2315 2320 2325 Gly Ser Arg His Gln Leu Gln Ser Ala Asp Ser Ser Arg His Ser 2330 2335 2340 Gly Ile Gly His Gly Gln Ala Ser Ser Ala Val Arg Asp Ser Gly 2345 2350 2355 His Arg Gly Ser Ser Gly Ser Gln Ala Ile Asp Ser Glu Gly His 2360 2365 2370 Ser Glu Asp Ser Asp Thr Gln Ser Val Ser Ala Gln Gly Gln Ala 2375 2380 2385 Gly Pro His Gln Arg Ser His Lys Glu Ser Ala Arg Gly Gln Ser 2390 2395 2400 Gly Glu Ser Ser Gly Arg Ser Gly Ser Phe Leu Tyr Gln Val Ser 2405 2410 2415 Thr His Glu Gln Pro Glu Ser Thr His Gly Gln Ser Ala Pro Ser 2420 2425 2430 Thr Gly Gly Arg Gln Gly Ser His His Asp Gln Ala Gln Asp Ser 2435 2440 2445 Ser Arg His Ser Ala Ser Gln Glu Gly Gln Asp Thr Ile Arg Gly 2450 2455 2460 His Pro Gly Pro Ser Arg Gly Gly Arg Gln Gly Ser His His Lys 2465 2470 2475 Gln Ser Val Asp Arg Ser Gly His Ser Gly Ser His His Ser His 2480 2485 2490 Thr Thr Ser Gln Gly Arg Ser Asp Ala Ser Arg Gly Gln Ser Gly 2495 2500 2505 Ser Arg Ser Ala Ser Arg Gln Thr His Asp Lys Glu Gln Ser Gly 2510 2515 2520 Asp Gly Ser Arg His Ser Gly Ser Arg His His Glu Ala Ser Ser 2525 2530 2535 Trp Ala Asp Ser Ser Arg His Ser Gln Ala Gly Gln Gly Gln Ser 2540 2545 2550 Glu Gly Ser Arg Thr Ser Arg Arg Gln Gly Ser Ser Phe Ser Gln 2555 2560 2565 Asp Ser Asp Ser Glu Gly His Ser Glu Asp Ser Glu Arg Arg Ser 2570 2575 2580 Gly Ser Ala Ser Arg Asn His Arg Gly Ser Ala Gln Glu Gln Ser 2585 2590 2595 Arg Asp Gly Ser Arg His Pro Arg Ser His His Glu Asp Arg Ala 2600 2605 2610 Gly His Gly His Ser Ala Asp Ser Ser Arg Gln Ser Gly Thr His 2615 2620 2625 His Ala Gln Asn Ser Ser Gly Gly Gln Ala Ala Ser Phe His Glu 2630 2635 2640 Gln Ala Arg Ser Ser Ala Gly Glu Arg His Gly Ser His His Gln 2645 2650 2655 Gln Ser Ala Asp Ser Ser Arg His Ser Gly Ile Gly His Gly Gln 2660 2665 2670 Ala Ser Ser Ala Val Arg Asp Ser Gly His Arg Gly Ser Ser Gly 2675 2680 2685 Ser Gln Ala Ser Asp Ser Glu Gly His Ser Glu Asp Ser Asp Thr 2690 2695 2700 Gln Ser Val Ser Ala His Gly Gln Ala Gly Pro His Gln Gln Ser 2705 2710 2715 His Gln Glu Ser Thr Arg Gly Arg Ser Ala Glu Arg Ser Gly Arg 2720 2725 2730 Ser Gly Ser Phe Leu Tyr Gln Val Ser Thr His Glu Gln Ser Glu 2735 2740 2745 Ser Ala His Gly Arg Thr Gly Pro Ser Thr Gly Gly Arg Gln Gly 2750 2755 2760 Ser Arg His Glu Gln Ala Arg Asp Ser Ser Arg His Ser Ala Ser 2765 2770 2775 Gln Glu Gly Gln Asp Thr Ile His Gly His Pro Gly Ser Arg Arg 2780 2785 2790 Gly Gly Arg Gln Gly Ser Tyr His Glu Gln Ser Val Asp Arg Ser 2795 2800 2805 Gly His Ser Gly Ser His His Ser His Thr Thr Ser Gln Gly Arg 2810 2815 2820 Ser Asp Ala Ser His Gly Gln Ser Gly Ser Arg Ser Ala Ser Arg 2825 2830 2835 Gln Thr Arg Asn Glu Gln Gln Ser Gly Asp Gly Ser Arg His Ser 2840 2845 2850 Gly Ser Ser His His Glu Ala Ser Thr Gln Ala Asp Ser Ser Arg 2855 2860 2865 His Ser Gln Ser Gly Gln Gly Glu Ser Ala Gly Ser Arg Arg Ser 2870 2875 2880 Arg Arg Gln Gly Ser Ser Val Ser Gln Asp Ser Asp Ser Glu Ala 2885 2890 2895 Tyr Pro Glu Asp Ser Glu Arg Arg Ser Glu Ser Ala Ser Arg Asn 2900 2905 2910 Arg His Gly Ser Ser Arg Glu Gln Ser Arg Asp Gly Ser Arg His 2915 2920 2925 Pro Gly Ser Ser His Arg Asp Thr Thr Arg His Val Gln Ser Ser 2930 2935 2940 Pro Val Gln Ser Asp Ser Ser Thr Ala Lys Glu His Gly His Phe 2945 2950 2955 Ser Ser Leu Ser Gln Asp Ser Ala Tyr Arg Ser Gly Ile Gln Ser 2960 2965 2970 Arg Gly Ser Pro His Ser Ser Ser Ser Tyr His Tyr Gln Ser Glu 2975 2980 2985 Gly Thr Glu Arg Gln Lys Gly Gln Ser Gly Leu Val Trp Arg His 2990 2995 3000 Gly Ser Tyr Gly Ser Ala Asp Tyr Asp Tyr Gly Glu Ser Gly Phe 3005 3010 3015 Arg His Ser Gln His Gly Ser Val Ser Tyr Asn Ser Asn Pro Val 3020 3025 3030 Val Phe Lys Glu Arg Ser Asp Ile Cys Lys Ala Ser Ala Phe Gly 3035 3040 3045 Lys Asp His Pro Arg Tyr Tyr Ala Thr Tyr Ile Asn Lys Asp Pro 3050 3055 3060 Gly Leu Cys Gly His Ser Ser Asp Ile Ser Lys Gln Leu Gly Phe 3065 3070 3075 Serl Gln Ser Gln Arg Tyr Tyr Tyr Glu 3080 3085 <210> 21 <211> 2764 <212> PRT <213> Pan troglodytes <400> 21 Met Ser Thr Leu Leu Glu Asn Ile Phe Ala Ile Ile Asn Leu Phe Lys 1 5 10 15 Gln Tyr Ser Lys Lys Asp Lys Asn Thr Asp Thr Leu Ser Ser Lys Glu 20 25 30 Leu Lys Glu Leu Glu Lys Glu Phe Arg Gln Ile Leu Lys Asn Pro 35 40 45 Asp Asp Pro Asp Met Val Asp Val Phe Met Asp His Leu Asp Ile Asp 50 55 60 His Asn Lys Lys Ile Asp Phe Thr Glu Phe Leu Leu Met Val Phe Lys 65 70 75 80 Leu Ala Gln Ala Tyr Glu Ser Thr Arg Lys Glu Asn Leu Pro Ile 85 90 95 Ser Gly His Lys His Arg Lys His Ser His His Asp Lys His Glu Asp 100 105 110 Asn Lys Gln Glu Glu Asn Lys Glu Asn Arg Lys Arg Pro Ser Ser Leu 115 120 125 Glu Arg Arg Asn Asn Arg Lys Gly Asn Lys Gly Arg Ser Lys Ser Pro 130 135 140 Arg Glu Thr Gly Gly Lys Arg His Glu Ser Ser Ser Glu Lys Lys Glu 145 150 155 160 Arg Lys Gly Tyr Ser Pro Thr His Arg Glu Glu Glu Tyr Gly Lys Asn 165 170 175 His His Asn Ser Ser Lys Lys Gln Lys Asn Lys Thr Glu Asn Thr Arg 180 185 190 Leu Glu Asp Asn Arg Lys Arg Leu Ser Glu Arg Leu Glu Glu Lys Glu 195 200 205 Asp Asn Glu Glu Gly Gly Tyr Asp Tyr Glu Asn Thr Gly Arg Met Thr 210 215 220 Gln Lys Trp Ile Gln Ser Gly His Ile Ala Thr Tyr Tyr Thr Ile Gln 225 230 235 240 Asp Glu Ala Tyr Asp Thr Thr Asp Asn Leu Leu Glu Glu Asn Lys Ile 245 250 255 Tyr Glu Arg Ser Arg Ser Ser Asp Asp Lys Ser Ser Ser Gln Val Asn 260 265 270 Arg Ser Arg His Glu Asn Thr Ser Gln Val Pro Leu Gln Glu Ser Arg 275 280 285 Thr Arg Lys Arg Arg Gly Ser Arg Val Ser Gln Asp Arg Asp Ser Glu 290 295 300 Gly His Ser Glu Asp Ser Glu Arg His Ser Gly Ser Ala Ser Arg Asn 305 310 315 320 His Pro Gly Ser Ala Arg Gln Gln Ser Arg Asp Gly Ser Arg His Pro 325 330 335 Arg Ser His Asp Glu Asp Arg Ala Ser His Gly His Ser Ala Asp Ser 340 345 350 Ser Arg Gln Ser Gly Thr Arg His Ala Glu Thr Ser Ser Arg Gly Gln 355 360 365 Thr Ala Ser Ser His Glu Gln Ala Arg Ser Ser Pro Gly Glu Arg His 370 375 380 Gly Ser Arg His Gln Gln Ser Ala Asp Ser Ser Arg His Ser Ala Thr 385 390 395 400 Gly Arg Gly Gln Ala Ser Ser Ala Val Ser Asp Arg Gly Pro Arg Gly 405 410 415 Ser Ser Gly Ser Gln Ala Ser Asp Ser Glu Gly His Ser Glu Asn Ser 420 425 430 Asp Thr Gln Ser Val Ser Gly His Gly Lys Ala Gly Pro Arg Gln Gln 435 440 445 Ser His Gln Glu Ser Ala Arg Gly Arg Ser Gly Glu Arg Ser Gly His 450 455 460 Ser Gly Ser Phe Leu Tyr Gln Val Ser Thr His Glu Gln Ser Asp Ser 465 470 475 480 Ala His Gly Arg Thr Gly Thr Ser Thr Gly Gly Arg Gln Gly Ser His 485 490 495 His Glu Gln Ala Arg Asp Ser Ser Arg His Ser Ala Ser Gln Glu Gly 500 505 510 Gln Asp Thr Ile Arg Gly His Pro Gly Ser Ser Arg Gly Gly Arg Gln 515 520 525 Gly Ser His His Glu Gln Leu Val Asn Arg Ser Gly His Ser Gly Ser 530 535 540 His His Ser His Thr Thr Ser Gln Gly Arg Ser Asp Ala Ser His Gly 545 550 555 560 Gln Ser Gly Ser Arg Ser Ala Ser Arg Gln Thr Arg Asn Glu Glu Gln 565 570 575 Ser Gly Asp Gly Ser Arg His Ser Gly Ser Arg His His Glu Ala Ser 580 585 590 Ser Gln Ala Asp Ser Ser Arg His Ser Gln Val Gly Gln Gly Gln Ser 595 600 605 Ser Gly Pro Arg Thr Ser Ser His Gln Gly Ser Ser Val Ser Gln Asp 610 615 620 Ser Asp Ser Glu Gly His Ser Glu Asp Ser Glu Arg Trp Ser Gly Ser 625 630 635 640 Ala Ser Arg Asn His His Gly Ser Ala Arg Glu Gln Ser Arg Asp Gly 645 650 655 Ser Arg His Pro Arg Ser His His Glu Asp Arg Ala Gly His Gly His 660 665 670 Ser Ala Asp Ser Ser Arg Gln Ser Gly Thr Arg His Thr Glu Ser Ser 675 680 685 Ser Arg Gly Gln Ala Val Ser Ser His Glu Gln Ala Arg Ser Ser Pro 690 695 700 Gly Glu Arg His Gly Ser Arg His Gln Gln Ser Ala Asp Ser Ser Arg 705 710 715 720 His Ser Gly Ile Gly His Gly Gln Ala Ser Ser Ala Val Arg Asp Ser 725 730 735 Gly His Arg Gly Ser Ser Gly Ser Gln Ala Ile Asp Ser Glu Gly His 740 745 750 Ser Glu Asp Ser Asp Thr Gln Ser Val Ser Ser His Gly Gln Ala Gly 755 760 765 Pro His Gln Gln Ser His Lys Glu Ser Ala Arg Gly Gln Ser Gly Glu 770 775 780 Ser Ser Gly Arg Ser Gly Ser Phe Leu Tyr Gln Val Ser Ser His Glu 785 790 795 800 Gln Ser Glu Ser Thr Tyr Gly Gln Thr Ala Pro Ser Thr Gly Gly Arg 805 810 815 Gln Gly Ser Arg His Glu Gln Ala Arg Asn Ser Ser Arg His Ser Ala 820 825 830 Ser Gln Asp Gly Gln Asp Thr Ile Arg Gly His Pro Gly Ser Ser Arg 835 840 845 Gly Gly Arg Gln Gly Ser Tyr His Glu Gln Ser Val Asp Arg Ser Gly 850 855 860 His Ser Gly Tyr His His Ser His Thr Thr Pro Gln Gly Arg Ser Asn 865 870 875 880 Ala Ser His Gly Gln Ser Gly Ser Arg Ser Ala Ser Arg Gln Thr Arg 885 890 895 Asn Glu Glu Gln Ser Gly Asp Gly Ser Arg His Ser Gly Ser Arg His 900 905 910 His Glu Ala Ser Ser Arg Ala Asp Ser Ser Arg His Ser Gln Val Gly 915 920 925 Gln Gly Gln Ser Ser Gly Pro Arg Thr Ser Arg Asn Gln Gly Ser Ser 930 935 940 Val Ser Gln Asp Ser Asp Ser Gln Gly His Ser Glu Asp Ser Glu Arg 945 950 955 960 Trp Ser Gly Ser Ala Ser Arg Asn His His Gly Ser Ala Trp Glu Gln 965 970 975 Ser Arg Asp Gly Ser Arg His Pro Gly Ser His Gln Glu Asp Arg Ala 980 985 990 Gly His Gly His Ser Ala Asp Ser Ser Arg Gln Ser Gly Thr His His 995 1000 1005 Thr Glu Ser Ser Ser Arg Gly Gln Ala Ala Ser Ser His Glu His 1010 1015 1020 Ala Arg Ser Ser Ala Gly Glu Arg His Gly Ser His His Gln Gln 1025 1030 1035 Ser Ala Asp Ser Ser Arg His Ser Gly Ile Gly His Gly Gln Ala 1040 1045 1050 Ser Ser Ala Val Arg Asp Ser Gly His Arg Gly Ser Ser Gly Ser 1055 1060 1065 Gln Ala Ser Asp Ser Glu Gly His Ser Glu Asp Ser Asp Thr Gln 1070 1075 1080 Ser Leu Ser Ala His Gly Gln Ala Gly Pro His Gln Gln Ser His 1085 1090 1095 Gln Glu Ser Thr Arg Gly Arg Ser Ala Glu Arg Ser Gly Arg Ser 1100 1105 1110 Gly Ser Phe Leu Tyr Gln Val Ser Thr His Glu Gln Ser Glu Ser 1115 1120 1125 Ala His Gly Arg Thr Gly Thr Ser Thr Gly Gly Arg Lys Arg Ser 1130 1135 1140 Leu His Glu Gln Ala Arg Asp Ser Ser Arg His Ser Val Ser Gln 1145 1150 1155 Glu Gly Gln Asp Thr Ile His Gly His Ala Gly Ser Ser Ser Gly 1160 1165 1170 Gly Arg Gln Gly Ser His Tyr Glu Gln Leu Val Asp Arg Ser Gly 1175 1180 1185 His Ser Gly Ser His His Ser His Thr Thr Ser Gln Gly Arg Ser 1190 1195 1200 Asp Ala Tyr His Gly Gln Ser Gly Ser Arg Ser Ala Ser Arg Gln 1205 1210 1215 Thr His Asn Asp Glu Gln Ser Gly Asp Gly Phe Arg His Ser Gly 1220 1225 1230 Ser His His His Glu Ala Ser Ser Arg Ala Asp Ser Ser Arg His 1235 1240 1245 Ser Gln Val Gly Gln Gly Gln Ser Glu Gly Pro Arg Thr Ser Arg 1250 1255 1260 His Arg Glu Ser Ser Val Ser Gln Asp Ser Asp Ser Glu Gly His 1265 1270 1275 Ser Glu Asp Ser Glu Arg Trp Ser Gly Ser Ala Ser Arg Asn His 1280 1285 1290 His Gly Ser Ala Arg Glu Gln Ser Arg Asp Gly Ser Arg His Pro 1295 1300 1305 Arg Ser His His Glu Asp Arg Ala Gly His Gly His Ser Ala Asp 1310 1315 1320 Ser Ser Arg Gln Ser Gly Thr Arg His Thr Gln Thr Ser Ser Gly 1325 1330 1335 Gly Gln Ala Ala Ser Ser His Glu Gln Gly Arg Ser Ser Ala Gly 1340 1345 1350 Glu Arg His Gly Ser Arg His Gln Gln Ser Ala Asp Ser Ser Arg 1355 1360 1365 His Ser Gly Ile Gly His Gly Gln Ala Ser Ser Ala Val Arg Asp 1370 1375 1380 Ser Gly His Arg Gly Tyr Ser Gly Ser Gln Ala Ser Asp Ser Glu 1385 1390 1395 Gly His Ser Glu Asp Ser Asp Thr Gln Ser Val Ser Ala His Gly 1400 1405 1410 Gln Ala Gly Ser His Gln Gln Ser His Gln Glu Ser Ala Arg Gly 1415 1420 1425 Arg Ser Gly Glu Arg Ser Gly Arg Ser Gly Ser Phe Leu Tyr Gln 1430 1435 1440 Val Ser Thr His Glu Gln Ser Glu Ser Ser His Gly Trp Thr Gly 1445 1450 1455 Pro Ser Thr Arg Gly Arg Gln Gly Ser Arg His Glu Gln Ala Gln 1460 1465 1470 Asp Ser Ser Arg His Ser Ala Ser Gln Glu Gly Gln Asp Thr Ile 1475 1480 1485 Arg Gly His Pro Gly Pro Ser Arg Gly Gly Arg Gln Gly Tyr His 1490 1495 1500 His Glu Gln Ser Val Asp Ser Ser Gly His Ser Gly Ser His His 1505 1510 1515 Ser His Ile Thr Ser Gln Gly Ser Ser His Ala Ser His Gly Gln 1520 1525 1530 Ser Gly Ser Arg Ser Ala Ser Arg Thr Thr Arg Asn Asp Glu Gln 1535 1540 1545 Ser Val Asp Gly Ser Arg His Ser Gly Ser Arg His His Glu Ala 1550 1555 1560 Ser Thr His Ala Asp Ile Ser Arg His Ser Gln Ala Gly Gln Gly 1565 1570 1575 Gln Ser Glu Gly Ser Arg Thr Ser Arg Arg Gln Gly Ser Ser Val 1580 1585 1590 Ser Gln Asp Ser Asp Ser Glu Gly His Ser Glu Asp Ser Glu Arg 1595 1600 1605 Trp Ser Gly Ser Ala Ser Arg Asn His Arg Gly Ser Ala Gln Glu 1610 1615 1620 Gln Ser Arg Asp Gly Ser Arg His Pro Arg Ser His His Glu Asp 1625 1630 1635 Arg Ala Gly His Gly His Ser Ala Asp Ser Ser Arg Gln Ser Gly 1640 1645 1650 Thr His His Ala Glu Thr Ser Ser Gly Gly Gln Ala Ala Ser Ser 1655 1660 1665 Arg Glu Gln Ala Arg Ser Ser Pro Gly Glu Arg His Gly Ser Arg 1670 1675 1680 His Gln Gln Ser Ala Asp Ser Ser Arg His Ser Gly Ile Arg Arg 1685 1690 1695 Gly Gln Ala Ser Ser Ala Val Arg Asp Ser Gly His Trp Gly Ser 1700 1705 1710 Ser Gly Ser Gln Ala Ser Asp Ser Glu Gly His Ser Glu Glu Ser 1715 1720 1725 Asp Thr Gln Ser Val Ser Gly His Gly Gln Asp Gly Pro His Gln 1730 1735 1740 Gln Ser His Gln Glu Ser Ala Arg Asp Arg Ser Gly Gly Arg Ser 1745 1750 1755 Gly Arg Ser Gly Ser Phe Leu Tyr Gln Val Ser Thr His Glu Gln 1760 1765 1770 Ser Glu Ser Thr His Gly Gln Thr Gly Thr Ser Thr Gly Gly Arg 1775 1780 1785 Gln Gly Ser His His Glu Gln Ala Arg Asp Ser Ser Arg His Ser 1790 1795 1800 Ala Ser Gln Glu Gly Gln Asp Thr Ile His Ala His Pro Gly Ser 1805 1810 1815 Ser Arg Gly Gly Arg Gln Gly Ser His His Glu Gln Ser Val Asp 1820 1825 1830 Thr Ser Gly His Ser Gly Ser His His Ser His Thr Thr Ser Gln 1835 1840 1845 Gly Arg Ser Asp Ala Ser His Gly Gln Ser Gly Ser Arg Ser Ala 1850 1855 1860 Ser Arg Gln Thr Arg Asn Asp Glu Gln Ser Gly Asp Gly Ser Arg 1865 1870 1875 His Ser Gly Ser His His His Glu Ala Phe Thr Gln Ala Asp Ser 1880 1885 1890 Ser Arg His Ser Gln Ser Gly Gln Gly Glu Ser Ala Gly Ser Arg 1895 1900 1905 Arg Ser Arg Arg Gln Gly Ser Ser Val Ser Gln Asp Ser Asp Ser 1910 1915 1920 Gln Gly His Ser Glu Asp Ser Glu Arg Trp Ser Gly Ser Ala Ser 1925 1930 1935 Arg Asn Gln His Gly Ser Ala Arg Glu Gln Ser Arg Asp Gly Ser 1940 1945 1950 Arg His Pro Gly Ser His Gln Glu Asp Arg Ala Gly His Gly His 1955 1960 1965 Ser Ala Asp Ser Ser Arg Gln Ser Gly Thr Arg His Thr Glu Ser 1970 1975 1980 Ser Ser Arg Gly Gln Ala Ala Ser Ser His Glu Gln Ala Arg Ser 1985 1990 1995 Ser Ala Gly Glu Arg His Gly Ser Arg His Gln Leu Gln Ser Ala 2000 2005 2010 Asp Ser Ser Arg His Ser Gly Ile Gly His Gly Gln Ala Ser Ser 2015 2020 2025 Ala Val Arg Asp Ser Gly His Arg Gly Ser Ser Gly Ser Gln Ala 2030 2035 2040 Ile Asp Ser Glu Gly His Ser Glu Asp Ser Asp Thr Gln Ser Val 2045 2050 2055 Ser Ala Gln Gly Gln Ala Gly Pro His Gln Arg Ser His Lys Glu 2060 2065 2070 Ser Ala Arg Gly Gln Ser Gly Glu Ser Ser Gly Arg Ser Gly Ser 2075 2080 2085 Phe Leu Tyr Gln Val Ser Thr His Glu Gln Pro Glu Ser Thr His 2090 2095 2100 Gly Gln Ser Ala Pro Ser Thr Gly Gly Arg Gln Gly Ser His His 2105 2110 2115 Asp Gln Ala Gln Asp Ser Ser Arg His Ser Ala Ser Gln Glu Gly 2120 2125 2130 Gln Asp Thr Ile Arg Gly His Pro Gly Pro Ser Arg Gly Gly Arg 2135 2140 2145 Gln Gly Ser His His Lys Gln Ser Val Asp Arg Ser Gly His Ser 2150 2155 2160 Gly Ser His His Ser His Thr Thr Ser Gln Gly Arg Ser Asp Ala 2165 2170 2175 Ser Arg Gly Gln Ser Gly Ser Arg Ser Ala Ser Arg Gln Thr His 2180 2185 2190 Asp Lys Glu Gln Ser Gly Asp Gly Ser Arg His Ser Gly Ser Arg 2195 2200 2205 His His Glu Ala Ser Ser Trp Ala Asp Ser Ser Arg His Ser Gln 2210 2215 2220 Ala Gly Gln Gly Gln Ser Glu Gly Ser Arg Thr Ser Arg Arg Gln 2225 2230 2235 Gly Ser Ser Phe Ser Gln Asp Ser Asp Ser Glu Gly His Ser Glu 2240 2245 2250 Asp Ser Glu Arg Arg Ser Gly Ser Ala Ser Arg Asn His Arg Gly 2255 2260 2265 Ser Ala Gln Glu Gln Ser Arg Asp Gly Ser Arg His Pro Arg Ser 2270 2275 2280 His His Glu Asp Arg Ala Gly His Gly His Ser Ala Asp Ser Ser 2285 2290 2295 Arg Gln Ser Gly Thr His His Ala Gln Asn Ser Ser Gly Gly Gln 2300 2305 2310 Ala Ala Ser Phe His Glu Gln Ala Arg Ser Ser Ala Gly Glu Arg 2315 2320 2325 His Gly Ser His His Gln Gln Ser Ala Asp Ser Ser Arg His Ser 2330 2335 2340 Gly Ile Gly His Gly Gln Ala Ser Ser Ala Val Arg Asp Ser Gly 2345 2350 2355 His Arg Gly Ser Ser Gly Ser Gln Ala Ser Asp Ser Glu Gly His 2360 2365 2370 Ser Glu Asp Ser Asp Thr Gln Ser Val Ser Ala His Gly Gln Ala 2375 2380 2385 Gly Pro His Gln Gln Ser His Gln Glu Ser Thr Arg Gly Arg Ser 2390 2395 2400 Ala Glu Arg Ser Gly Arg Ser Gly Ser Phe Leu Tyr Gln Val Ser 2405 2410 2415 Thr His Glu Gln Ser Glu Ser Ala His Gly Arg Thr Gly Pro Ser 2420 2425 2430 Thr Gly Gly Arg Gln Gly Ser Arg His Glu Gln Ala Arg Asp Ser 2435 2440 2445 Ser Arg His Ser Ala Ser Gln Glu Gly Gln Asp Thr Ile His Gly 2450 2455 2460 His Pro Gly Ser Arg Arg Gly Gly Arg Gln Gly Ser Tyr His Glu 2465 2470 2475 Gln Ser Val Asp Arg Ser Gly His Ser Gly Ser His His Ser His 2480 2485 2490 Thr Thr Ser Gln Gly Arg Ser Asp Ala Ser His Gly Gln Ser Gly 2495 2500 2505 Ser Arg Ser Ala Ser Arg Gln Thr Arg Asn Glu Gln Gln Ser Gly 2510 2515 2520 Asp Gly Ser Arg His Ser Gly Ser Ser His His Glu Ala Ser Thr 2525 2530 2535 Gln Ala Asp Ser Ser Arg His Ser Gln Ser Gly Gln Gly Glu Ser 2540 2545 2550 Ala Gly Ser Arg Arg Ser Arg Arg Gln Gly Ser Ser Val Ser Gln 2555 2560 2565 Asp Ser Asp Ser Glu Ala Tyr Pro Glu Asp Ser Glu Arg Arg Ser 2570 2575 2580 Glu Ser Ala Ser Arg Asn Arg His Gly Ser Ser Arg Glu Gln Ser 2585 2590 2595 Arg Asp Gly Ser Arg His Pro Gly Ser Ser His Arg Asp Thr Thr 2600 2605 2610 Arg His Val Gln Ser Ser Pro Val Gln Ser Asp Ser Ser Thr Ala 2615 2620 2625 Lys Glu His Gly His Phe Ser Ser Leu Ser Gln Asp Ser Ala Tyr 2630 2635 2640 Arg Ser Gly Ile Gln Ser Arg Gly Ser Pro His Ser Ser Ser Ser 2645 2650 2655 Tyr His Tyr Gln Ser Glu Gly Thr Glu Arg Gln Lys Gly Gln Ser 2660 2665 2670 Gly Leu Val Trp Arg His Gly Ser Tyr Gly Ser Ala Asp Tyr Asp 2675 2680 2685 Tyr Gly Glu Ser Gly Phe Arg His Ser Gln His Gly Ser Val Ser 2690 2695 2700 Tyr Asn Ser Asn Pro Val Val Phe Lys Glu Arg Ser Asp Ile Cys 2705 2710 2715 Lys Ala Ser Ala Phe Gly Lys Asp His Pro Arg Tyr Tyr Ala Thr 2720 2725 2730 Tyr Ile Asn Lys Asp Pro Gly Leu Cys Gly His Ser Ser Asp Ile 2735 2740 2745 Ser Lys Gln Leu Gly Phe Ser Gln Ser Gln Arg Tyr Tyr Tyr Tyr 2750 2755 2760 Glu <210> 22 <211> 819 <212> PRT <213> Bos taurus <400> 22 Met Ser Thr Leu Leu Glu Asn Ile Asn Asp Ile Ile Lys Ile Phe His 1 5 10 15 Lys Tyr Ser Lys Thr Asp Lys Glu Thr Asp Thr Leu Ser Glu Lys Glu 20 25 30 Leu Lys Glu Leu Val Glu Val Glu Phe Arg Pro Ile Leu Lys Asn Pro 35 40 45 Gly Asp Pro Asp Thr Ala Glu Val Phe Met Tyr Asn Leu Asp Arg Asp 50 55 60 His Asn Asn Lys Ile Asp Phe Thr Glu Phe Phe Leu Met Val Phe Lys 65 70 75 80 Val Ala Gln Val Tyr Tyr Ser Tyr Thr Gln Arg Gln Asn Leu Gln Arg 85 90 95 Ala Gly Gln Lys Gln Lys Lys Cys Thr Tyr His Tyr Gly Asp Glu Glu 100 105 110 Asp Asp Thr Glu Glu Asp Lys Glu Glu Thr Glu Arg Lys Tyr Ser His 115 120 125 Ser Arg Ser Asp Gly Lys Thr Gln Asp Arg Ser Lys Ser Pro Arg Gly 130 135 140 Arg Gly Lys Lys Arg His Gly Ser Lys Ser Gly Ser Lys Gln Arg Arg 145 150 155 160 Gly Asp Thr Pro Thr Ser Gly Leu Arg His Gly Cys Ser Lys Lys His 165 170 175 Glu Ser Arg Arg Glu Lys Lys Arg Arg Pro Ser Ser Thr Glu Pro Lys 180 185 190 Glu Arg Arg His Met Ser Ser Val Ser Pro Thr Arg Gly Tyr Glu Glu 195 200 205 Lys Glu Glu Glu His Gly Tyr Glu Asn Lys Gly Lys Thr Ser Ala Lys 210 215 220 Cys Ile Gly Ser Glu Tyr Asp Asp Ser Tyr Gln Val Cys Glu Asp Lys 225 230 235 240 Val Thr Thr Asn Phe Gln Pro Ser His Ser Lys Asn Tyr Gly Ser Asn 245 250 255 Ile Thr Lys Gly Arg Asp Thr Glu Gly His Ser Arg Asp Thr Gly Arg 260 265 270 Lys Ser Val Phe Thr His Ala Arg Ser Gly Ser Ser Ser Arg Asn Gln 275 280 285 Asn Gly Ser Val Gln Thr His Thr Gly Asp Asn Ser Thr His Ser Glu 290 295 300 Ser Gln Gln Glu Thr Asn Ser Glu Ser Val His Arg Arg Ser Arg Asn 305 310 315 320 Thr Gly Gln Arg Gln Gly Ser His His Glu Gln Ser Arg Asp Ser Ser 325 330 335 Arg His Ser Gly Thr Arg His Gly Gln Pro Ser Thr Gly Ser Gly Gly 340 345 350 Ser Arg His Arg Glu Ser Ser Val Ser Gln Ala Ser Asp Ser Glu Gly 355 360 365 His Ser Glu Asp Ser Gly Arg Gln Ser Val Thr Thr His Gly Arg Pro 370 375 380 Gly Ser Ser Ser Arg Asn Gln His Gly Ser Ser Gln Gly Gln Thr Gly 385 390 395 400 Asp Ser Ser Arg His Ser Glu Ser His Gln Gly Arg His Ser Asp Ser 405 410 415 Leu Gln Arg Arg Ser Gly Thr Ser Thr Gly Gln Arg Gln Gly Ser His 420 425 430 His Glu Gln Ser Arg Asp Ser Ser Arg His Ser Gly Thr Gln Gln Gly 435 440 445 Gln Thr Ser Thr Gly Ser Gly Ser Ser Arg His Arg Asp Ser Ser Val 450 455 460 Ser Gln Ala Ser Asp Ser Glu Gly His Ser Glu Asp Ser Gly Arg Gln 465 470 475 480 Leu Glu Thr Val Leu Gly Ile Gln Ser Pro Ile Lys Glu Ala Thr Val 485 490 495 Lys Leu Phe Met Lys Gly Gln Asp Pro Ala Arg Gly Lys Gly Arg Gly 500 505 510 Ala Thr Met Ser Ser Gln Gly Thr Ala Pro Asp Thr Leu Glu Leu Asp 515 520 525 Met Asp Lys Ser Gln Leu Asn Leu Glu Met Ala Asp Ile Gly Asn Pro 530 535 540 Val Leu Val Lys Pro Val Thr Val Arg Asp Thr Pro Glu Ile Gln Val 545 550 555 560 Gly Ile Leu Arg Gln Leu Met Glu Asp Met Val Pro Ala Gln Gly Thr 565 570 575 Asn Met Asp Pro Pro Met Val Arg Gln Gly Thr Val Ile Gly Thr Gln 580 585 590 Arg Pro Ile Lys Gly Asp Thr Ile Arg Asp Thr Gln Glu Ile Gln Val 595 600 605 Gly Ile Leu Arg Gln Leu Met Glu Asp Leu Val Pro Ala Gln Glu Thr 610 615 620 Asn Met Asp Leu Pro Met Ala Arg Val Glu Thr Val Leu Gly Thr Gln 625 630 635 640 Ser Pro Ile Lys Arg Gly Ile Val Asn Leu Ser With Glu Gly Gln Asp 645 650 655 Pro Ala Leu Gly Lys Asp Arg Gly Ala Thr With Ser Ser Gln Glu Thr 660 665 670 Ala Pro Asp Thr Leu Glu Leu Asp With Gly Lys Pro Gln Leu Asn Leu 675 680 685 Glu Met Ala Asp Ile Gly Asn Pro Val Leu Val Lys Pro Val Thr Val 690 695 700 Arg Asp Thr Pro Glu Ile Gln Val Gly Ile Leu Arg Gln Leu With Glu 705 710 715 720 Gly Leu Val Gln Ala Gln Glu Thr Asn With Asp Leu Pro Arg Ala Arg 725 730 735 Gln Glu Thr Val Leu Gly Thr Gln His Ile Ile Lys Gly Gly Thr Val 740 745 750 Asn Leu Ser Met Lys Gly Gln Arg Leu Glu Leu Gly Lys Asp Arg Gly 755 760 765 Ala Thr Met Ser Ser Gln Gly Thr Ala Pro Asp Ser Leu Glu Leu Asp 770 775 780 Met Asp Ile Pro Leu Leu Asp Leu Glu Met Ala Asp Ile Gly Asn Pro 785 790 795 800 Val Leu Val Ser Gln Val Thr Glu Arg Asp Thr Gln Glu Ile Gln Val 805 810 815 Asp Ser Pro <210> 23 <211> 1183 <212> PRT <213> Macaca mulatta <400> 23 Met Ser Thr Leu Leu Glu Asn Ile Phe Ala Ile Ile Asn Leu Phe Lys 1 5 10 15 Gln Tyr Ser Lys Lys Asp Lys Asn Thr Asp Thr Leu Ser Lys Lys Glu 20 25 30 Leu Lys Glu Leu Leu Glu Lys Glu Phe Arg Pro Ile Leu Lys Asn Pro 35 40 45 Asp Asp Pro Asp Thr Val Glu Val Phe Met Asp His Leu Asp Ile Asp 50 55 60 His Asn Lys Lys Ile Asp Phe Thr Glu Phe Leu Leu Met Val Phe Lys 65 70 75 80 Leu Ala Gln Ala Tyr Tyr Glu Ser Thr Arg Lys Gln Asn Leu Pro Ile 85 90 95 Ala Gly His Lys His Arg Lys His Ser His His Asp Lys His Glu Asp 100 105 110 Asn Lys Glu Glu Glu Asn Lys Glu Lys Arg Lys Arg Pro Leu Ser Leu 115 120 125 Glu Arg Arg Asn Asn Arg Lys Gly Asn Thr Gly Arg Ser Lys Ser Pro 130 135 140 Arg Glu Arg Gly Gly Lys Arg His Glu Ser Ser Phe Glu Lys Lys Glu 145 150 155 160 Arg Lys Gly Tyr Ser Pro Thr Tyr Glu Glu Glu Glu Tyr Gly Gln Asn 165 170 175 His His Lys Ser Ser Lys Lys Glu Lys Asn Lys Thr Glu Ile Thr Arg 180 185 190 Leu Glu His Glu Gly Lys Arg Ile Ser Glu Arg Pro Glu Lys Lys Glu 195 200 205 Glu Lys Glu Asp Gly Gln Phe Asp Tyr Glu Asn Ala Gly Arg Met Asp 210 215 220 Glu Lys Trp Thr Glu Ser Gly His Ile Ala Ile Tyr His Ala Ile Gln 225 230 235 240 Asp Glu Val Asp Asp Thr Thr Glu Asn Ile Leu Glu Glu Asn Arg Arg 245 250 255 Tyr Glu Thr Ser Arg Ser Pro His Asp Lys Ser Ser Leu Arg Val Asn 260 265 270 Arg Ser Pro Asn Ala Asn Thr Ser Gln Ile Pro Leu Val Glu Pro Arg 275 280 285 Arg Arg Thr Arg Gln Arg Ser Ser Val Ser Gln Asp Ser Asp Ser Glu 290 295 300 Gly His Ser Glu Asp Ser Glu Arg Gln Ser Glu Ser Ala Ser Arg Asn 305 310 315 320 His His Gly Ser Val Arg Glu Gln Ser Arg His Gly Ser Arg His Pro 325 330 335 Gly Ser His His Glu Asp Arg Ala Gly His Gly His Ser Ala Asp Ser 340 345 350 Ser Thr Gln Ser Gly Thr Arg His Thr Glu Thr Ser Ser Arg Gly Gln 355 360 365 Ala Val Ser Ser His Glu Gln Ala Arg Ser Ser Pro Gly Glu Arg His 370 375 380 Gly Ser Arg His Gln Gln Ser Ala Glu Ser Ser Arg His Ser Gly Ile 385 390 395 400 Gly Arg Gly Gln Ala Ser Ser Ala Val Ser Asp Arg Gly His Gln Gly 405 410 415 Pro Ser Gly Ser His Phe Ser Asp Ser Glu Gly His Ser Glu His Ser 420 425 430 Asp Thr Gln Ser Val Ser Gly His Gly Gln Ala Gly Pro His Pro His 435 440 445 Ser His Gln Glu Ser Ala Arg Gly Arg Ser Arg Glu Arg Ser Gly Arg 450 455 460 Ser Gly Ser Phe Leu Tyr Gln Val Ser Thr His Glu Gln Ser Glu Ser 465 470 475 480 Thr His Gly Arg Thr Gly Pro Ser Ser Ala Gly Arg Gln Gly Ser Arg 485 490 495 Asn Glu Gln Ala Arg Asp Ser Ser Arg His Ser Ala Ser His Glu Val 500 505 510 Gln Asp Thr Val His Gly His His Gly Ser Ser Arg Gly Arg Arg Gln 515 520 525 Gly Ser His His Glu Gln Leu Val Asp Ser Ser Gly His Ser Gly Ser 530 535 540 His His Ser His Thr Thr Ser Gln Gly Arg Ser Asp Ala Ser Arg Gly 545 550 555 560 Glu Ser Gly Ala Arg Ser Ala Ser Arg Gln Thr Arg His Glu Glu Gln 565 570 575 Ser Gly Asp Gly Ser Arg His Ser Gly Ser Arg His His Glu Ser Ser 580 585 590 Asn Arg Ala Asp Ser Ser Arg His Ala Gln Ser Ser Gln Gly Gln Ser 595 600 605 Ala Gly Phe Arg Thr Ser Thr Arg Arg Gly Ser Ser Val Ser Gln Asp 610 615 620 Ser Asp Ser Glu Gly His Ser Glu Asp Ser Glu Arg Gln Ser Glu Ser 625 630 635 640 Ala Ser Arg Asn His His Gly Ser Val Arg Glu Gln Ser Arg His Gly 645 650 655 Ser Gly His Ser Gly Ser His His Gln Asp Lys Val Gly His Arg Tyr 660 665 670 Ser Gly Asp Ser Ser Arg Gln Ser Gly Thr His His Val Glu Thr Ser 675 680 685 Ser His Gly Gln Ala Ala Ser Ser His Glu Gln Thr Arg Ser Ser Pro 690 695 700 Gly Glu Arg His Gly Ser His His Gln Gln Ser Ala Asp Ser Ser Arg 705 710 715 720 His Ser Gly Thr Gly Arg Gly Gln Ala Ser Ser Ala Val Ser Asp Arg 725 730 735 Gly His Gln Gly Pro Ser Gly Ser His Phe Ser Asp Ser Glu Gly His 740 745 750 Ser Glu His Ser Asp Thr Gln Ser Val Ser Gly Gln Gly Gln Ala Gly 755 760 765 Arg His Pro His Ser His Gln Glu Ser Ala Arg Gly Arg Ser Gly Glu 770 775 780 Arg Ser Gly Arg Ser Gly Ser Phe Val Tyr Gln Val Ser Thr His Glu 785 790 795 800 Gln Ser Glu Ser Thr His Gly Arg Thr Gly Pro Ser Thr Gly Gly Arg 805 810 815 Gln Gly Ser Arg Asn Glu Gln Ala Arg Asp Ser Ser Arg His Ser Val 820 825 830 Ser His Glu Gly Gln Asp Thr Ile His Gly His His Gly Ser Ser Arg 835 840 845 Gly Gly Arg Gln Gly Ser His His Glu Gln Ser Val Asp Ser Ser Gly 850 855 860 His Ser Gly Ser His His Ser His Thr Thr Ser Gln Gly Arg Ser Asp 865 870 875 880 Ala Ser Arg Gly Glu Ser Gly Ala Arg Ser Ala Ser Arg Gln Thr Arg 885 890 895 His Glu Glu Gln Ser Gly Asp Gly Ser Arg His Ser Gly Ser Arg His 900 905 910 His Glu Ala Ser Asn Arg Ala Asp Ser Ser Arg His Ala Gln Ser Gly 915 920 925 Gln Gly Gln Ser Ala Gly Phe Arg Thr Ser Thr Arg Arg Gly Ser Ser 930 935 940 Val Ser Gln Asp Ser Asp Ser Glu Gly His Ser Glu Asp Ser Glu Arg 945 950 955 960 Gln Ser Glu Ser Ala Ser Arg Asn His His Gly Ser Val Arg Glu Gln 965 970 975 Ser Arg His Gly Ser Arg His Pro Gly Ser His His Glu Asp Arg Ala 980 985 990 Gly His Gly His Ser Ala Asp Ser Ser Arg Gln Ser Gly Thr Arg His 995 1000 1005 Thr Glu Thr Ser Ser Arg Gly Gln Ala Val Ser Ser His Glu Gln 1010 1015 1020 Ala Arg Ser Ser Pro Gly Glu Arg His Gly Ser Arg His Gln Gln 1025 1030 1035 Ser Ala Glu Ser Ser Arg His Ser Gly Ile Gly Arg Gly Gln Ala 1040 1045 1050 Ser Ser Ala Val Ser Asp Arg Gly His Gln Gly Pro Ser Gly Ser 1055 1060 1065 His Phe Ser Asp Ser Glu Gly His Ser Glu His Ser Asp Thr Gln 1070 1075 1080 Ser Val Ser Gly His Gly Gln Ala Gly Pro His Pro His Ser His 1085 1090 1095 Gln Glu Ser Ala Arg Gly Arg Ser Gly Glu Arg Ser Gly Arg Ser 1100 1105 1110 Gly Ser Phe Leu Tyr Gln Val Ser Thr His Glu Gln Ser Glu Ser 1115 1120 1125 Thr His Gly Arg Thr Gly Pro Ser Ser Ala Gly Arg Gln Gly Ser 1130 1135 1140 Arg Asn Glu Gln Ala Arg Asp Ser Ser Arg His Ser Ala Ser His 1145 1150 1155 Glu Val Gln Asp Thr Val His Gly His His Gly Ser Ser Arg Gly 1160 1165 1170 Gly Arg Gln Gly Ser His His Glu Gln Ser 1175 1180 <210> 24 <211> 2265 <212> PRT <213> Macaca mulatta <400> 24 Met Thr Asp Leu Leu Arg Ser Val Val Thr Val Ile Asp Val Phe Tyr 1 5 10 15 Lys Tyr Thr Lys Gln Asp Gly Glu Cys Gly Thr Leu Ser Lys Asp Glu 20 25 30 Leu Lys Glu Leu Leu Glu Lys Glu Phe His Pro Val Leu Lys Asn Pro 35 40 45 Asp Asp Pro Asp Thr Val Asp Val Ile Met His Met Leu Asp Arg Asp 50 55 60 His Asp Arg Arg Leu Asp Phe Thr Glu Phe Leu Leu Met Ile Phe Lys 65 70 75 80 Leu Thr Met Ala Cys Asn Lys Val Leu Ser Lys Glu Tyr Cys Lys Ala 85 90 95 Ser Gly Ser Lys Lys His Arg Arg Gly His Arg His Gln Glu Glu Glu 100 105 110 Ser Glu Thr Glu Glu Asp Glu Glu Asp Thr Pro Glu His Lys Ser Gly 115 120 125 Tyr Arg His Ser Ser Trp Ser Glu Gly Glu Glu His Gly Tyr Ser Ser 130 135 140 Gly His Ser Arg Gly Thr Val Lys Arg Arg His Gly Ser Asn Ser Arg 145 150 155 160 Arg Leu Gly Arg Gln Gly His Leu Ser Ser Ser Gly Asn Gln Glu Arg 165 170 175 Ser Gln Lys Arg Tyr His Arg Ser Ser Ser Gly His Ser Trp Ser Ser 180 185 190 Gly Lys Glu Arg His Gly Phe Ser Ser Gly Glu Leu Arg Glu Arg Ile 195 200 205 Asn Lys Ser His Val Ser Pro Ser Arg Glu Phe Gly Glu Glu Tyr Glu 210 215 220 Ser Gly Ser Gly Ser Lys Ser Trp Glu Arg Lys Gly His Gly Gly Leu 225 230 235 240 Ser Cys Gly Leu Glu Ile Ser Gly His Glu Ser Asn Ser Thr Gln Ser 245 250 255 Arg Ser Ser Gly Gln Lys Leu Gly Ser Ser Arg Ser Cys Ser Gly Asp 260 265 270 Ser Arg Arg Arg Ser His Ala Cys Gly Tyr Ser Asn Ser Ser Gly Cys 275 280 285 Gly Arg Pro Gln Asn Ala Ser Asn Ser Cys Gln Ser His Arg Phe Gly 290 295 300 Gly Gln Val Asn Gln Ser Ser Tyr Ile Gln Ser Gly Cys Gln Ser Gly 305 310 315 320 Ile Asn Gly Glu Gln Gly His Asp Cys Val Ser Gly Gly Gln Pro Ser 325 330 335 Gly Cys Gly Gln Pro Glu Ser Asn Ser Cys Ser Gln Ser Tyr Ser Gln 340 345 350 Arg Gly Tyr Gly Ala Arg Glu Asn Gly Gln Pro Gln Asn Cys Gly Gly 355 360 365 Gln Gln Arg Thr Gly Ser Ser Gln Ser Ser Phe Cys Gly Gln Tyr Glu 370 375 380 Ser Gly Gly Ser Gln Ser Cys Ser Asn Gly Gln His Glu His Gly Ser 385 390 395 400 Cys Gly Arg Phe Ser Asn Ser Ser Ser Ser Asn Glu Phe Ser Lys Cys 405 410 415 Gly Lys His Arg Ser Gly Ser Gly Gln Phe Thr Ser Cys Glu Gln His 420 425 430 Gly Thr Gly Leu Ser Gln Ser Ser Gly Phe Glu Gln Gln Val Ala Gly 435 440 445 Ser Ser Gln Thr Cys Ser Gln Tyr Gly Ser Arg Ser Ser Gln Ser Ser 450 455 460 Gly Tyr Asp Glu His Gly Ser Ser Ser Gly Lys Thr Ser Gly Phe Gly 465 470 475 480 Gln His Arg Ser Gly Ser Gly His Ser Ser Gly Phe Gly Gln His Gly 485 490 495 Ser Gly Ser Gly Gln Ser Phe Gly Phe Gly Gln His Gly Ser Gly Ser 500 505 510 Gly Gln Ser Ser Gly Phe Gly Gln His Glu Ser Arg Ser Cys Gln Ser 515 520 525 Ser Tyr Gly Gln His Gly Ser Gly Ser Ser Gln Ser Ser Gly Tyr Gly 530 535 540 Gln His Ala Ser Arg Gln Thr Ser Gly Phe Gly Gln His Gly Leu Gly 545 550 555 560 Ser Gly Gln Ser Thr Gly Phe Gly Gln Tyr Gly Ser Gly Ser Gly Gln 565 570 575 Ser Ser Gly Phe Gly Gln His Gly Ser Gly Ser Gly Gln Ser Ser Gly 580 585 590 Phe Gly Gln His Glu Ser Arg Ser Gly Gln Ser Ser Tyr Gly Gln His 595 600 605 Ser Ser Ser Ser Ser Gln Ser Ser Gly Tyr Gly Gln His Gly Ser Arg 610 615 620 Gln Thr Ser Gly Phe Gly Gln His Gly Ser Gly Ser Gly Gln Ser Thr 625 630 635 640 Gly Phe Gly Gln Tyr Gly Ser Ser Leu Gly Gln Ser Ser Gly Phe Gly 645 650 655 Gln His Gly Ser Gly Ser Gly Gln Ser Ser Gly Phe Gly Gln His Glu 660 665 670 Ser Thr Ser Gly Gln Ser Ser Tyr Gly Gln His Gly Phe Gly Ser Ser 675 680 685 Gln Ser Ser Gly Cys Gly Gln His Gly Leu Ser Ser Gly Gln Thr Ser 690 695 700 Gly Phe Gly Gln His Glu Leu Ser Ser Gly Gln Ser Ser Ser Phe Gly 705 710 715 720 Gln His Gly Ser Gly Ser Gly Gln Ser Ser Gly Phe Arg Gln His Glu 725 730 735 Ser Gly Ser Gly Gln Ser Ser Gly Phe Gly Gln His Glu Ser Arg Ser 740 745 750 His Gln Ser Ser Tyr Gly Pro His Gly Ser Gly Ser Gly Gln Ser Ser 755 760 765 Gly Tyr Gly Gln His Gly Ser Ser Ser Gly Gln Thr Ser Gly Phe Gly 770 775 780 Gln Gln Gly Ser Ser Ser Ser Gln Tyr Ser Gly Phe Gly Gln His Gly 785 790 795 800 Ser Gly Leu Gly Gln Ser Ser Gly Phe Gly Gln His Gly Thr Gly Ser 805 810 815 Gly Gln Phe Ser Gly Phe Gly Gln His Glu Ser Arg Ser His Gln Ser 820 825 830 Ser Tyr Gly Gln His Gly Ser Gly Ser Ser Gln Ser Ser Gly Tyr Gly 835 840 845 Gln His Gly Ser Ser Ser Gly His Thr Thr Gly Phe Gly Gln His Arg 850 855 860 Ser Ser Ser Gly Gln Tyr Ser Gly Phe Gly Gln His Gly Ser Gly Leu 865 870 875 880 Gly Gln Ser Ser Gly Phe Gly Gln His Gly Thr Gly Ser Gly Gln Ser 885 890 895 Ser Gly Phe Gly Gln His Glu Ser Arg Ser His Gln Ser Ser Tyr Gly 900 905 910 Gln His Gly Ser Gly Ser Ser Gln Ser Ser Ser Tyr Gly Gln His Gly 915 920 925 Ser Ser Ser Gly Gln Thr Ser Gly Phe Gly Gln His Arg Ser Gly Ser 930 935 940 Gly Gln Ser Ser Gly Phe Gly Gln Tyr Gly Leu Gly Ser Gly Gln Ser 945 950 955 960 Ser Gly Phe Gly Gln His Gly Ser Gly Thr Gly Gln Ser Ser Gly Phe 965 970 975 Ala Arg His Glu Tyr Arg Ser Gly Gln Ser Ser Tyr Gly Gln His Gly 980 985 990 Thr Gly Ser Ser Gln Ser Ser Gly Cys Gly Gln Arg Glu Ser Gly Ser 995 1000 1005 Gly Pro Thr Thr Gly Phe Gly Gln His Val Ser Gly Ser Asp Asn 1010 1015 1020 Phe Ser Ser Ser Gly Gln His Ile Ser Gly Ser Asp Gln Ser Thr 1025 1030 1035 Gly Phe Gly Gln Tyr Gly Ser Gly Ser Gly Gln Ser Thr Gly Leu 1040 1045 1050 Gly Gln Val Glu Ser Gln Gln Val Ala Ser Gly Ser Thr Val His 1055 1060 1065 Gly Arg Gln Glu Thr Thr His Gly Gln Thr Ile Asn Thr Ala Arg 1070 1075 1080 His Ser Gln Ser Gly Gln Gly Gln Ser Thr Gln Thr Gly Ser Arg 1085 1090 1095 Val Ser Arg Arg Arg Arg Ser Ser Gln Ser Glu Asn Ile Asp Ser 1100 1105 1110 Glu Val His Ser Arg Val Ser His Arg His Ser Glu His Ile Asp 1115 1120 1125 Thr Gln Val Gly Ser His Tyr Pro Glu Ser Gly Ser Thr Val His 1130 1135 1140 Arg Arg Gln Gly Thr Thr His Gly Gln Arg Gly Asp Thr Thr Arg 1145 1150 1155 His Ser His Ser Gly His Gly Gln Ser Thr Gln Thr Gly Ser Arg 1160 1165 1170 Thr Thr Gly Arg Gln Arg Phe Ser His Ser Asp Ala Thr Asp Ser 1175 1180 1185 Glu Val His Ser Gly Val Ser His Arg Pro His Ser Gln Glu His 1190 1195 1200 Thr His Gly Gln Asp Gly Ser Gln Leu Gly Glu Ser Gln Ser Thr 1205 1210 1215 Val His Glu Arg His Glu Thr Thr Tyr Gly Gln Thr Gly Asp Ala 1220 1225 1230 Thr Gly His Gly Tyr Ser Gly His Gly Gln Ser Thr Gln Ile Gly 1235 1240 1245 Ser Arg Thr Ser Gly Arg Arg Gly Ser Gly His Ser Glu Ser Ser 1250 1255 1260 Asp Thr Glu Val His Ser Gly Gly Ser His Arg Pro His Ser Gln 1265 1270 1275 Glu Gln Thr His Gly Gln Ala Arg Ser Gln His Gly Glu Ser Arg 1280 1285 1290 Ser Thr Val His Glu Arg His Gly Thr Thr His Gly Gln Thr Gly 1295 1300 1305 Asp Thr Thr Arg Tyr Ala His Tyr His Asn Gly Gln Ser Ala Gln 1310 1315 1320 Arg Gly Ser Arg Thr Thr Gly Arg Gly Ser Gly His Ser Glu Tyr 1325 1330 1335 Ser Asp Ser Glu Leu Tyr Ser Gly Gly Ser His Thr Tyr Ser Gly 1340 1345 1350 His Thr His Gly Gln Ala Gly Ser Gln His Gly Glu Ser Asp Ser 1355 1360 1365 Ile Val His Glu Arg Tyr Gly Thr Thr His Gly Gln Thr Gly Asp 1370 1375 1380 Thr Thr Arg His Ala His Tyr Ser His Gly Gln Ser Lys Gln Arg 1385 1390 1395 Gly Ser Arg Thr Thr Gly Arg Arg Gly Ser Gly His Ser Glu Tyr 1400 1405 1410 Ser Asp Ser Glu Gly His Ser Gly Gly Ser His Thr His Ser Gly 1415 1420 1425 His Thr His Gly His Thr His Gly Gln Ala Gly Ser Gln His Gly 1430 1435 1440 Glu Ser Gly Ser Ser Gly His Gly Gly Gln Gly Thr Thr His Gly 1445 1450 1455 Gln Thr Gly Asp Thr Thr Arg His Ala His Tyr Gly His Gly Gln 1460 1465 1470 Ser Thr Gln Arg Gly Ser Arg Thr Ala Gly Arg Arg Gly Ser Gly 1475 1480 1485 His Ser Glu Tyr Ser Asp Ser Glu Gly His Ser Gly Val Ser His 1490 1495 1500 Thr His Ser Gly His Thr His Gly Gln Ala Gly Ser Gln His Gly 1505 1510 1515 Glu Ser Glu Ser Thr Val His Glu Arg Gln Gln Thr Thr His Gly 1520 1525 1530 Gln Thr Gly Asp Thr Thr Arg His Ala His Tyr Gly His Gly Gln 1535 1540 1545 Ser Thr Gln Thr Gly Ser Arg Thr Thr Gly Arg Arg Gly Ser Gly 1550 1555 1560 His Ser Glu Tyr Ser Asp Ser Glu Gly His Ser Gly Val Ser His 1565 1570 1575 Thr His Ser Gly His Thr His Gly Gln Ala Arg Ser Gln His Gly 1580 1585 1590 Glu Ser Gly Ser Ala Ile His Gly Arg Gln Gly Thr Ile His Gly 1595 1600 1605 Gln Thr Gly Asp Thr Thr Arg His Gly Gln Ser Gly His Gly Gln 1610 1615 1620 Ser Thr Gln Thr Gly Ser Arg Thr Thr Gly Arg Arg Gly Ser Gly 1625 1630 1635 His Ser Glu Tyr Ser Asp Ser Glu Gly His Ser Gly Gly Ser His 1640 1645 1650 Thr His Ser Gly His Thr His Gly Gln Ala Gly Ser Gln His Gly 1655 1660 1665 Glu Ser Gly Ser Thr Val His Gly Arg Gln Gly Thr Ile His Gly 1670 1675 1680 Gln Thr Gly Asp Thr Thr Arg His Gly Gln Ser Gly His Gly Gln 1685 1690 1695 Ser Ile Glu Thr Gly Ser Arg Thr Thr Gly Arg Arg Gly Ser Gly 1700 1705 1710 His Ser Glu T...

Claims

1. A recombinant microorganism capable of secreting a filaggrin (FLG) polypeptide, comprising an expression vector comprising a first coding sequence comprising a gene capable of expressing the filaggrin polypeptide and a second coding sequence comprising a gene capable of expressing a cell-penetrating peptide, wherein the filaggrin polypeptide consists of FLG domains 9-10 (FLG domain[9-10]); the FLG domain 9-10 (FLG domain [9-10]) has at least 90% sequence identity with SEQ ID NO: 46; The recombinant microorganism is selected from the group consisting of Bifidobacterium, Brevibacterium, Propionibacterium, Lactococcus, Streptococcus, Staphylococcus, Lactobacillus, Enterococcus, Pediococcus, Leuconostoc, or Oenococcus, and combinations thereof. The recombinant microorganism.

2. The recombinant microorganism of claim 1, further comprising a third coding sequence comprising a gene capable of expressing an export signal.

3. The recombinant microorganism of claim 2, wherein expression of the first coding sequence, the second coding sequence and the third coding sequence is under the control of a promoter.

4. The recombinant microorganism of claim 3 , wherein the first coding sequence, the second coding sequence and the third coding sequence are arranged in frame.

5. 3. The recombinant microorganism of claim 2, wherein the first coding sequence, the second coding sequence, and the third coding sequence are operably linked to a promoter.

6. The recombinant microorganism of claim 2 , wherein the recombinant microorganism is a bacterium or a combination of bacteria.

7. The recombinant microorganism of claim 1, wherein the filaggrin (FLG) polypeptide consists of an amino acid sequence selected from SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, or SEQ ID NO:

6.

8. 2. The recombinant microorganism of claim 1, wherein the recombinant microorganism is Staphylococcus epidermidis.

9. The recombinant microorganism of claim 1 , wherein the microorganism secretes a filaggrin fusion protein.

10. 1. A method for producing a live biotherapeutic composition, comprising: (a) transfecting a cell with (i) a first coding sequence comprising a nucleic acid sequence capable of expressing a therapeutic polypeptide, wherein the therapeutic polypeptide consists of an FLG domain 9-10 (FLG domain [9-10]) polypeptide, wherein the FLG domain 9-10 (FLG domain [9-10]) polypeptide has at least 90% sequence identity to SEQ ID NO: 46, and (ii) a second coding sequence comprising a nucleic acid sequence capable of expressing a cell-penetrating peptide; (b) causing the transfected cells to produce the therapeutic polypeptide fusion protein; (c) obtaining a live biotherapeutic composition; Including, The recombinant microorganism is selected from the group consisting of Bifidobacterium, Brevibacterium, Propionibacterium, Lactococcus, Streptococcus, Staphylococcus, Lactobacillus, Enterococcus, Pediococcus, Leuconostoc, or Oenococcus, and combinations thereof. The method.

11. 11. The method of claim 10, further comprising (iii) transfecting the cell with a third coding sequence comprising a nucleic acid sequence capable of expressing a localization signal.

12. 12. The method of claim 11, wherein the first coding sequence, the second coding sequence and the third coding sequence are located in a single plasmid.

13. 12. The method of claim 11, wherein the arrangement of the first coding sequence, the second coding sequence and the third coding sequence is operably linked to a promoter.

14. 11. The method of claim 10, wherein the cell is Staphylococcus epidermidis.

15. A composition obtained by the method according to any one of claims 10 to 14.

16. 16. The composition of claim 15, comprising a pharmaceutically acceptable carrier selected from the group consisting of an aqueous solution, an emulsion, a cream, a lotion, a gel, or an ointment.

17. 1. A live biotherapeutic composition comprising a recombinant microorganism, the recombinant microorganism comprising: (i) a first coding sequence comprising a nucleic acid sequence capable of expressing a therapeutic polypeptide, wherein the therapeutic polypeptide consists of a filaggrin domain 9-10 (FLG domain [9-10]) polypeptide, wherein the filaggrin domain 9-10 (FLG domain [9-10]) polypeptide has at least 90% sequence identity to SEQ ID NO: 46; (ii) a second coding sequence comprising a nucleic acid sequence capable of expressing a cell-penetrating peptide; (iii) a third coding sequence comprising a nucleic acid sequence capable of expressing a localization signal; and (iv) a promoter operably linked to the first coding sequence, the second coding sequence, and the third coding sequence. Including, the first coding sequence, the second coding sequence, and the third coding sequence are capable of expressing a filaggrin fusion product; The recombinant microorganism is selected from the group consisting of Bifidobacterium, Brevibacterium, Propionibacterium, Lactococcus, Streptococcus, Staphylococcus, Lactobacillus, Enterococcus, Pediococcus, Leuconostoc, or Oenococcus, and combinations thereof. The live biotherapeutic composition.

18. 18. The composition of claim 17, wherein the recombinant microorganism is Staphylococcus epidermidis.

19. 19. The composition of claim 17 or 18, wherein the export signal exports the filaggrin fusion product, or a variant thereof, out of the recombinant microorganism.

20. 19. The composition of claim 17 or 18, wherein the cell-penetrating peptide facilitates entry of the filaggrin fusion product, or a variant thereof, into human keratinocytes.

21. 18. The composition of claim 17, comprising a pharmaceutically acceptable carrier selected from the group consisting of an aqueous solution, an emulsion, a cream, a lotion, a gel, or an ointment.

22. A kit comprising the composition of any one of claims 17 to 21 and instructions for use.

23. A composition for treating a skin disease in a subject, comprising a recombinant microorganism according to any one of claims 1 to 9, or a composition according to any one of claims 15 to 21, or a kit according to claim 22.

24. 24. The composition of claim 23, wherein the skin disease is atopic dermatitis.

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