45 results about "Toxin protein" patented technology

The invention provides a production strain of an antibiotic-free plasmid, the production strain is a gene editing strain of a PIR strain and is named as PIR1-WN:: 0636 or PIR1-PR: 0636, the production strain contains a nucleotide sequence for coding toxin protein and the antibiotic-free plasmid, and the antibiotic-free plasmid contains a nucleotide sequence for coding antitoxin protein; and preferably, the replicon DNA element of the nonreactive plasmid is R6K-gamma. The toxin protein gene of the production strain disclosed by the invention can be stably passaged, has lethality after being induced and can be used for plasmid screening; according to the invention, the positive rate of transforming the nonreactive plasmid into the PIR1-WN:: 0636 strain is more than 80%, and stable production of the plasmid with a high superhelix ratio can be realized.

The invention belongs to the technical field of target sample characterization, and provides application of enterococcus pore-forming toxin mutant protein in nanopore detection. The enterococcus pore-forming toxin mutant protein is EPX1 nanopore mutant protein and is obtained by mutating wild type EPX1 nanopore protein, the amino acid sequence of the wild type EPX1 nanopore protein is as shown in SEQ ID NO: 1, and the mutation means that one or more amino acids of the wild type EPX1 nanopore protein are mutated into common amino acids except original amino acids. The EPX1 protein (wild type) is a type of pore-forming toxin protein and widely exists in various living bodies in nature, and a mutant of the EPX1 protein shows excellent pore-forming capacity after being modified (after amino acid mutation). The structural stability, the molecular recognition efficiency and the detection performance of the EPX1 nanopore mutant protein are remarkably improved.

The invention provides a predatory mite venom protein or a spider venom protein. The amino acid sequences of the predatory mite venom protein and the spider venom protein are selected from SEQ ID NO.1-8 or any one of SEQ ID NO.1-8 with 95% or more of identity. Tetranychus urticae tests show that after spraying treatment, the predatory mite poison protein or spider poison protein provided by the invention achieves a 100% lethal effect within 48 hours, and is a known protein pesticide with the highest tetranychus urticae lethal rate at present. The predatory mite poison protein or spider poison protein provided by the invention has high specificity to spider mites, is mild and harmless to human bodies and other organisms in the environment, can be used in farmlands, gardens and even family potted plants, and can be naturally degraded; the production process is simple and efficient, and has the potential of large-scale application.

A biopesticide composition and a method of making the composition. The biopesticide composition comprises a biopesticide in the form of an endotoxin protein, preferably a δ-endotoxin; and a late embryogenesis abundant (LEA) protein in the form of a LEA2 / dehydrin protein, wherein the ratio of endotoxin to LEA2 / dehydrin proteins is 1:40. The LEA2 / dehydrin proteins in the composition confers heat resistance to the endotoxin up to a temperature of about 42° C. The method of making the biopesticide composition comprises providing a biopesticide, providing a LEA protein, by isolating genes for a LEA protein from a plant and expressing the isolated gene for the LEA protein in an expression vector to produce a recombinant LEA protein, the method including combining the biopesticide and the LEA protein.

The application provides a method for establishing a skin injury model of ICR mice and application thereof; ICR mice are used as experimental animals, and a local skin bump is formed by injecting a mucor toxin protein on the back of the near-tail region to form a skin injury model; the model can be applied in mucor infection research, and research related to mucor toxin protein in mucormycosis can be quickly and conveniently carried out, and a new method is provided for basic research and clinical application related to mucormycosis infection and mucor toxin protein.

The application provides application of an intermediodorsal or spider ovigorous granule toxin-VI protein in preparation of an anti-inflammatory reagent, which can inhibit M1 type macrophage activation, promote M2 type macrophage cell transformation, inhibit macrophage and microglia cell activation and inhibit LPS-induced inflammation. The intermediodorsal or spider ovigorous granule toxin-VI protein disclosed by the application can significantly inhibit excessive activation of inflammation in an LPS-induced inflammation model, has good anti-inflammatory effects on peripheral inflammation and neural inflammation, has good anti-inflammatory effects in in-vivo and in-vitro inflammation models, and experiments show that the intermediodorsal or spider ovigorous granule toxin-VI protein has no obvious toxic side effects on mice at an inflammation inhibiting dose, and has good clinical application prospects.

Disclosed is an OTA analysis method based on a high-efficiency homogeneous electrochemiluminescence aptamer sensor, which comprises the following steps: preparing a mixed solution of L / S*-Ru, IA / B, hairpin probe H1, hairpin probe H2 and exonuclease Exo-III; putting OTA into the mixed solution for reaction to obtain a reaction solution; pouring the reaction solution into a solution containing 0.02M TPrA to obtain a test solution; and immersing an electrode in the test solution for ECL detection and reading the detection result. The analysis method has the advantages of high stability, high sensitivity and good selectivity, and can be used for detecting other toxins, proteins and other biomolecules by changing the aptamer sequence and blocking chain.

This invention belongs to the field of biotechnology and discloses a monoclonal antibody that specifically binds to the recombinant protein of Clostridium perfringens (Cp) β2 toxin, a hybridoma cell line, and its applications. This invention provides a monoclonal antibody that specifically binds to the β2 toxin protein and a hybridoma cell line CPB2-2A4 (accession number CCTCC NO: C2024312) that secretes this monoclonal antibody. The monoclonal antibody reacts only with the Cp β2 toxin protein and does not cross-react with other pathogens infecting cattle and sheep, exhibiting extremely high specificity. The kit utilizes competitive ELISA technology, using a mouse monoclonal antibody that specifically binds to the Cp β2 toxin recombinant protein as a competing antibody, enabling the detection of Cp β2 toxin antibodies in serum from various animal sources, avoiding species limitations of sample sources. Furthermore, the kit exhibits good reproducibility. The kit has relatively relaxed technical requirements during operation and can be widely used in production for applications such as Cp epidemiological surveys and vaccine immunization level monitoring.

The present disclosure is directed, in part, to saxiphilin proteins, nucleic acids encoding the same, compositions comprising the same, kits comprising the same, and methods of detecting toxin in samples. In some embodiments, the toxin is saxitoxin or derivatives of the same.

The invention provides a Chinese beetle pyemotes skin necrosis toxin gene as well as an encoding protein and application thereof, and belongs to the technical field of biology. The toxin gene provided by the invention comprises the following nucleotide sequences: (a) a nucleotide sequence as shown in SEQ ID NO: 1; (b) a nucleotide sequence as shown in SEQ ID NO: 2; (c) a nucleotide sequence as shown in SEQ ID NO: 3; (d) a nucleotide sequence as shown in SEQ ID NO: 4; and (e) a nucleotide sequence having at least 95% sequence consistency with at least one of the nucleotide sequences as shown in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4. The pyemotes sinensis toxin protein provided by the invention is derived from natural toxin protein and has good insecticidal activity on spodoptera frugiperda.

The invention discloses a recombinant tetanus toxin protein rTT as well as a preparation method and application thereof, the recombinant protein sequentially contains a His tag, a modified exogenous polypeptide and two tetanus toxin fragments connected in series through flexible polypeptides from the N end to the C end, the two tetanus toxin fragments connected in series form two intramolecular disulfide bonds in a molecule through double mutation, and the tetanus toxin fragments are connected in series through flexible polypeptides. Coding nucleotide of the recombinant protein is expressed by escherichia coli BL21 (DE3), induced at low temperature and purified, and target protein with the purity larger than or equal to 95% can be obtained. A subunit vaccine prepared from the protein and a two-way oil adjuvant ISA201 enables mice to achieve 100% protection under 100 LD50 challenge dose, the protection effect is better than that of truncated tetanus toxin heavy chain fragment TTc protein, and the recombinant protein can be used for tetanus prevention vaccines and serological diagnostic reagents.

The present invention relates to an antitoxin protein binding molecule and a kit thereof, the antitoxin protein binding molecule comprises an antitoxin protein targeting antibody or an antigen binding fragment thereof, the binding molecule comprises a light chain variable region and a heavy chain variable region, the light chain variable region comprises a complementarity determining region LCDR, and the heavy chain variable region comprises a heavy chain variable region LCDR. The LCDR comprises an LCDR1 of which the sequence is SEQ ID NO: 1, an LCDR2 of which the sequence is SEQ ID NO: 2 and an LCDR3 of which the sequence is SEQ ID NO: 3; the heavy chain variable region comprises a complementarity determining region HCDR, and the HCDR comprises an HCDR1 of which the sequence is SEQ ID NO: 4, an HCDR2 of which the sequence is SEQ ID NO: 5 and an HCDR3 of which the sequence is SEQ ID NO: 6.

The invention provides a Hainan needle scorpion active polypeptide and application thereof. The Hainan double-needle scorpion active polypeptide is derived from Hainan double-needle scorpion toxin protein, and the amino acid sequence of the Hainan double-needle scorpion active polypeptide is shown as SEQ ID NO.1. The full length of the polypeptide toxin is 38 amino acid residues, the polypeptide toxin comprises 6 cysteines, the whole polypeptide is slightly alkaline, the isoelectric point is 11.87, the hydrophilic amino acid residues account for more than 50%, and the theoretical molecular weight is 4200.78 Da. The difference between the actual molecular weight and the theoretical molecular weight is 7Da, which shows that six cysteines in the polypeptide form three pairs of disulfide bonds. The Hainan needle scorpion active polypeptide disclosed by the invention has the activity of selectively inhibiting a voltage-gated potassium ion channel Kv1.3, so that the Hainan needle scorpion active polypeptide can be applied to preparation of polypeptide medicines for treating autoimmune diseases.

The invention provides three Cry1Ac modified toxin proteins with high insecticidal activity and application of the Cry1Ac modified toxin proteins, and relates to the field of genetic engineering and biological control. The amino acid sequences of the three modified proteins are sequentially shown as SEQ ID NO: 1, SEQ ID NO: 3 and SEQ ID NO: 5, and the amino acid sequences are sequentially shown as SEQ ID NO: 2, SEQ ID NO: 4 and SEQ ID NO: 6. The insect-resistant protein is a Cry1Ac toxin modified protein with improved insecticidal activity, which is screened from a Cry1Ac toxin key region saturation mutation library targeting Asiatic corn borer cadherin and ABC transporter, has higher insecticidal activity on lepidoptera pests, can be used for preventing and controlling agricultural pests and resisting the Bt resistance problem, and has important scientific and practical significance.

This invention relates to an antitoxin protein binding molecule and a kit thereof. The antitoxin protein binding molecule comprises an antibody or an antigen-binding fragment targeting the antitoxin protein. The binding molecule includes a light chain variable region and a heavy chain variable region. The light chain variable region includes a complementarity-determining region (LCDR), which comprises LCDR1 with the sequence SEQ ID NO:1, LCDR2 with the sequence SEQ ID NO:2, and LCDR3 with the sequence SEQ ID NO:3. The heavy chain variable region includes a complementarity-determining region (HCDR), which comprises HCDR1 with the sequence SEQ ID NO:4, HCDR2 with the sequence SEQ ID NO:5, and HCDR3 with the sequence SEQ ID NO:6.

There is provided an antibiotic-free microplasmid comprising a nucleotide sequence encoding an antitoxin protein and a replicon. The skeleton of the non-antibiotic microplasmid is small and is controlled within 1000bp, redundant useless fragments are reduced, a resistance expression cassette is not contained, the utilization rate of a target sequence in the plasmid is improved, and the production burden is reduced. The method has higher efficiency and safety in plasmid DNA-mediated non-viral vector delivery, gene therapy, vaccine immunization, virus production, antibody production and other genetic engineering aspects. The invention also provides a system for producing plasmids based on a toxin-antitoxin system, the bacterial host cell contains a gene expression cassette capable of inducing expression of toxin protein, and the non-antibiotic microplasmids contain a gene expression cassette for expressing antitoxin protein and are used for efficiently maintaining replication and amplification of the non-antibiotic microplasmids.

The present invention discloses an HSPA8 protein that is resistant to duck Tembusu virus and its application. The present invention constructs a prokaryotic expression vector for expressing HSPA8, optimizes protein expression conditions, and expresses and purifies soluble, endotoxin-free HSPA8 proteins, including HSPA8-His protein and GST-HSPA8 protein. In vitro antiviral experiments demonstrate that the HSPA8 protein of the present invention has a good antiviral effect; in vivo antiviral experiments demonstrate that the HSPA8 protein can significantly reduce the pathogenicity of DTMUV infection, prevent viral proliferation in vivo, reduce weight loss in mice caused by viral infection, and reduce mortality in mice caused by viral infection. Therefore, the HSPA8 protein of the present invention has significant resistance to DTMUV infection and can be used to prepare drugs against duck Tembusu virus, providing a new strategy for the prevention and control of duck Tembusu virus.

The present invention relates to a method of purifying a non-toxic protein-removed Clostridium botulinum neurotoxin protein.

The invention provides a composition containing enterotoxin C. The composition comprises the following components: 5-100 ng / mL of enterotoxin C; 0.1 to 5 mg / mL of protein, wherein the protein is selected from whey protein, soy protein isolate or collagen; the concentration of the polypeptide is 0.05 to 3 mg / mL, and the polypeptide is collagen peptide or whey peptide with the molecular weight of 500 to 5000 Da; 1.0 to 8.0 mg / mL of an amino acid mixture, wherein the amino acid mixture comprises 18 essential and non-essential amino acids. The enterotoxin C-containing composition disclosed by the invention realizes remarkable breakthrough in the aspects of stability, targeting property and multifunctionality through a multi-component synergistic effect and technical innovation.

The present invention relates to a recombinant insecticidal gene for protecting agricultural crops and a method for protection from insect pests by modifying Chimeric δ-endotoxin protein Cry1EC to provide mCry1EC gene having SEQ ID NO.2 for a broader insecticidal host range and expressing the peptide in crop plants. The modification includes mutations at positions 475,476, and 483 in Domain III, wherein the amino acid Leucine, Glycine, and Lysine are replaced with Valine, Tryptophan, and Threonine respectively. The transgenic crop plant causes insect mortality upon feeding and renders protection from the plurality of insect pests.

Disclosed herein are pesticidal proteins that exhibit toxic activity against lepidoptera pest species, and that include, but are not limited to, TIC8643 and related pesticidal proteins. DNA constructs encoding the disclosed pesticidal proteins are provided. Transgenic plants, plant cells, seeds and plant parts that are resistant to lepidoptera insect infestation are provided, which contain recombinant nucleic acid sequences encoding insecticidal proteins of the invention, and vectors containing at least one coding sequence for expressing and delivering the encoded toxin proteins are also described. Also provided are methods for detecting the presence of a recombinant nucleic acid sequence or protein of the invention in a biological sample, as well as methods of using TIC8643 and related pesticidal proteins to control pests of Lepidoptera species.

The present application relates to the application of Staphylococcus aureus toxin protein related gene SA1833 / SA1832 in treating Staphylococcus aureus infection. Specifically, it relates to the application of Staphylococcus aureus toxin protein related gene SA1833 and / or SA1832 as a target in the preparation of a drug for treating drug-resistant Staphylococcus aureus infection, preferably, the drug resistance is preferably methicillin resistance. With SA1833 and / or SA1832 as a target, the clinical treatment of repeated and persistent infection caused by methicillin-resistant Staphylococcus aureus retention has a significant effect. It provides a new target for the development of drugs for treating repeated infection caused by clinical MRSA strains, provides more choices for the treatment of Staphylococcus aureus, and has a wide application prospect.

The application belongs to the technical field of characterizing target samples, and provides application of enterococcus pore-forming toxin mutant protein in nanopore detection. The enterococcus pore-forming toxin mutant protein is an EPX1 nanopore mutant protein, which is obtained by mutation of wild-type EPX1 nanopore protein, and the amino acid sequence of the wild-type EPX1 nanopore protein is shown as SEQ ID NO: 1. The mutation refers to that one or more amino acids of the wild-type EPX1 nanopore protein are mutated into common amino acids other than the original amino acids. The EPX1 protein (wild type) of the application is a kind of pore-forming toxin protein, which widely exists in various organisms in nature. The mutant thereof exhibits excellent pore-forming ability after modification (after amino acid mutation). The EPX1 nanopore mutant protein realizes significant improvement in structural stability, molecular recognition efficiency and detection performance.

Provided is an injectable vaccine comprising: a heat treated purified whole cell extract of B bronchiseptica, wherein this extract has been heated for 10 to 60 minutes at 45°C to 65°C; and p68 pertactin protein. Methods of making and methods of using the vaccine are also provided.

An expression vector for producing a target protein-fused pentamer of the present invention includes a polynucleotide that encodes a pentameric toxin protein and a viral nucleocapsid as fusion partners of a target protein. In addition, a method of producing a target protein-fused pentamer of the present invention includes manufacturing an expression vector for producing a target protein-fused pentamer, which includes a polynucleotide that encodes a target protein and a pentameric toxin protein and a viral nucleocapsid as fusion partners of the target protein; producing a transformant by introducing the expression vector for producing a target protein-fused pentamer into host cells; and culturing the transformant.

The invention provides a cell-free toxin synthesis system and method. The system at least comprises: (1) a cell-free system; (2) an amplification system containing amplified plasmids of the cell-free system is added, the plasmids are plasmids containing coding toxin protein, the amplification system containing the amplified plasmids is obtained after the plasmids are added into the amplification system according to the preset concentration to be amplified, the preset concentration is 2-10 ng / mu L, and the preset concentration is not 2 ng / mu L. According to the method, plasmids with a certain concentration are added into a cell-free reaction system, so that the in-vitro expression efficiency of toxins is effectively improved, and particularly, the purification efficiency is more obviously improved.

The invention discloses construction of an antibiotic-free plasmid stabilization system and application of the antibiotic-free plasmid stabilization system in riboflavin high-yield strains, and belongs to the field of gene engineering. The toxin protein YonT with a strong killing effect on escherichia coli K-12MG1655 is obtained through screening, the toxin protein YonT is subjected to successful inducible expression in an escherichia coli genome through toxin modification, and a constitutive repressor is expressed by plasmids, so that the inhibition effect on the modified toxin is achieved, and the effect of maintaining the stability of the plasmids is achieved. After 10 days of passage of a modified strain containing the system, the maintenance rate of plasmids is 84.9%. Escherichia coli containing the system is used as a chassis cell to express riboflavin, so that the constructed recombinant bacteria keep good plasmid stability in a 5L bioreactor and can produce riboflavin with the concentration of 11.5 g / L. The invention provides a novel plasmid stabilizing system, which is beneficial to expanding the application of engineering strains in industry.

The embodiment of the invention discloses a tRNA (transfer ribonucleic acid) site specific fragmentation-sequencing method based on gamma-toxin, and relates to the technical field of molecular biology. The technical problem that in the prior art, the recognition accuracy of a modification site of tRNA is low is solved. The tRNA site specific fragmentation-sequencing method based on gamma-toxin comprises the following steps: constructing to obtain a recombinant gamma-toxin protein, and purifying the recombinant gamma-toxin protein to obtain an intermediate product; carrying out a gamma-CT-seq process on the intermediate product; the gamma-CT-seq process comprises the following steps: enriching and demethylating small RNA (Ribonucleic Acid), demethylating AlkB in vitro, specifically cutting a gamma-toxin site, and constructing a library and sequencing.

The application belongs to the technical field of genetic engineering, and particularly relates to a single-domain antibody targeting Vibrio pathogenic PirA toxin protein and application thereof in immunodetection. The single-domain antibody targeting Vibrio pathogenic PirA toxin protein screened in the application is A02 or A15, the amino acid sequence of the single-domain antibody A02 is shown as SEQ ID NO. 1, and the amino acid sequence of the single-domain antibody A15 is shown as SEQ ID NO. 2. The single-domain antibody provided in the application is high in stability, strong in stress resistance, and good in specificity, and has good application value. The single-domain antibody provided in the application can be well applied to preparation of a detection product for detection of a shrimp AHPND pathogen.

The present invention relates to a method for purifying Clostridium botulinum toxin, comprising separating a non-toxin protein and an uncomplexed botulinum toxin protein, and performing a purification process of hydrophobic interaction and ion-exchange chromatography, thereby obtaining the uncomplexed botulinum toxin protein having high purity and increased yield. By using the botulinum purification method of the present invention, various industrial uses are possible and, in particular, botulinum toxin, which is in high demand in the pharmaceutical field, can be economically and highly-efficiently mass-produced, and thus can be effectively used in the industry.