73 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 polypeptide is used for preparing neurotoxin protein through recombinant expression and comprises a first restriction enzyme cutting site, a second restriction enzyme cutting site and a tag peptide fragment. The tag peptide fragment is used for affinity purification, the first restriction enzyme cutting site at least comprises one amino acid and is located on one side of the N end of the tag peptide fragment, and the second restriction enzyme cutting site at least comprises one amino acid and is located on one side of the C end of the tag peptide fragment. It is verified that after the polypeptide is used for fusion expression of neurotoxin, a large amount of neurotoxin target protein with biological activity can be rapidly prepared.

Disclosed herein are nucleotide sequences encoding an insecticidal protein exhibiting Lepidopteran inhibitory activity, as well as novel insecticidal proteins referred to herein as a BCW 001, BCW 002, BCW 003, and BCW toxic protein-containing chimeras and BCW toxin insecticide, transgenic plants expressing the chimeras or the insecticide, and methods for detecting the presence of the nucleotide sequences or the insecticide in a biological sample.

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 present invention relates to the technical field of agrochemicals, and in particular to highly-virulent metarhizium robertsii specifically modified for acrididae pests and a preparation method therefor and a use thereof. A protein comprises: a protein as shown in SEQ ID NO: 1, and / or a fusion protein thereof, and / or a homologous protein thereof. A toxin protein or a coding gene thereof is a key virulence factor of metarhizium acridum causing high virulence to locusts, and is only toxic to the acrididae pests and does not affect other insects. A recombinant entomopathogenic fungus effectively solves the disadvantage of original broad-spectrum entomopathogenic fungi having insufficient virulence to acrididae pests, and is also highly virulent to the acrididae pests while effectively controlling and preventing various other pests, thereby improving the pest prevention and control efficiency, achieving the effects of multi-purpose use with a single fungus and use cost saving, and not causing food and ecological safety issues.

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 present invention discloses a detection kit and a detection method for type B botulinum neurotoxin. The present invention can complete qualitative detection of various suspected cases of botulinum neurotoxin poisoning in a short time, provide evidence support for assisting doctors in diagnosis and timely treatment decisions, and is of great significance in social health and safety. At the same time, the detection method of the present invention can significantly improve the specificity of toxin proteins, avoid the time problems caused by animal experiments such as serotype identification, and can be applied to the research and development and production process of botulinum neurotoxin products. In addition, the present invention can perform quality control on the stability and shelf life of botulinum neurotoxin standards.

The invention belongs to the technical field of bioengineering, and discloses a monoclonal antibody specifically combined with clostridium septicum alpha toxin recombinant protein, a hybridoma cell strain and application. The invention provides a monoclonal antibody which is specifically combined with clostridium septicum alpha toxin protein and a hybridoma cell strain clostridium septicum alpha toxin-8H2 which secretes the monoclonal antibody and has the preservation number of CCTCC NO: C202535, the monoclonal antibody only reacts with the clostridium septicum alpha toxin protein, and does not have cross reaction with other pathogens which infect cattle and sheep; the specificity is extremely high; according to the kit, a competitive ELISA technology is utilized, a mouse monoclonal antibody specifically combined with clostridium septicum alpha toxin recombinant protein is taken as a competitive antibody, serum antibodies of various animal sources can be detected, and species limitation of sample sources is avoided. Besides, the kit is good in repeatability, relatively loose in technical requirements during operation, capable of being widely popularized and used in production, and applied to clostridium septicum epidemiological investigation and vaccine immune level monitoring.

The invention relates to a base editor DGhBE4max, a base editing method and application of the base editor DGhBE4max, and belongs to the technical field of gene editing. The invention provides a base editor DGhBE4max. The amino acid sequence of the base editor DGhBE4max is as shown in SEQ ID NO. 1; the base editor DGhBE4max comprises a deaminase rAPOBEC1, an endotoxin protein dDddA, an nCas9 protein, and a uracil glycosylase inhibitor UGI (Uracil Glucosylase Inhibitor). According to the base editor DGhBE4max, the editing efficiency of an existing cytosine base editor in plants can be remarkably improved, meanwhile, a base editing window can be remarkably expanded, and therefore a stable and efficient technical tool is provided for research of plant functional genes and directed evolution of key genes.

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 helicobacter pylori bispecific antibody comprises a first antibody, a connector and a second antibody, the first antibody (VHH) is specifically combined with a helicobacter pylori urease B subunit (UreB) antigen, the second antibody (ScFv) is specifically combined with a helicobacter pylori vacuolar toxin protein (VacA) antigen, the first antibody (VHH) is connected with the second antibody (ScFv) through the connector, and the second antibody (ScFv) is connected with the second antibody (ScFv) through the connector. The whole or part of the connector is a heavy chain constant region derived from humanized lgG1 or lgG2 or lgG3 or lgG4; the bispecific antibody can significantly reduce adhesion of helicobacter pylori in the stomach, and reduce infiltration of inflammatory cells and damage of stomach tissues; the helicobacter pylori bispecific antibody can quickly and obviously reduce the infection ability of helicobacter pylori, and does not generate other harmful effects on stomach tissues and cells.

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 invention relates to the technical field of bioengineering, in particular to recombinant botulinum toxin and a preparation method thereof. The nucleotide sequence of the coding gene of the recombinant botulinum neurotoxin is as shown in SEQ ID NO: 1; the amino acid sequence of the toxin is as shown in SEQ ID NO: 2. The clostridium botulinum neurotoxin sequence optimization method realizes soluble expression of clostridium botulinum neurotoxin protein by escherichia coli; meanwhile, the purity of the botulinum neurotoxin protein is improved by a novel purification process technology, and the double-chain botulinum neurotoxin with relatively high biological activity is obtained; according to the method, the process operation steps of the botulinum neurotoxin are reduced, the biological safety risk is remarkably reduced, industrial production amplification of the botulinum neurotoxin is facilitated, a solid foundation is provided for large-scale production of the botulinum neurotoxin, and the method can be effectively applied to preparation of medical, beauty and health-care products.

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.

The invention belongs to the technical field of biology, and discloses a monoclonal antibody specifically combined with clostridium perfringens (C.p) beta 2 toxin recombinant protein, a hybridoma cell strain and application. The invention provides a monoclonal antibody which is specifically combined with beta 2 toxin protein and a hybridoma cell strain CPB2-2A4 which secretes the monoclonal antibody and has the preservation number of CCTCC NO: C2024312, the monoclonal antibody only reacts with the C.p beta 2 toxin protein and does not have cross reaction with other pathogens which infect cattle and sheep, and the specificity is extremely high; according to the kit, a competitive ELISA technology is utilized, a mouse monoclonal antibody specifically combined with C.p beta2 toxin recombinant protein is taken as a competitive antibody, C.p beta2 toxin antibodies in serum of various animal sources can be detected, and species limitation of sample sources is avoided. In addition, the kit is good in repeatability. The kit has low technical requirements during operation, can be widely popularized and used in production, and is applied to C.p epidemiological investigation and vaccine immune level monitoring.

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 specifically relates to a genetically modified Yersinia entomophila engineered bacterium CSLH88-pCHSW and its application. The strain is classified and named as Yersinia entomophila engineered bacterium Yersinia entomophagous , which was deposited in the General Microbiological Center of China Microbiological Culture Collection Administration on August 6, 2021, with the deposit number CGMCC NO.23127. The Yersinia entomophila engineered bacteria CSLH88‑pCHSW obtained by the present invention can stably nptII-cry3AaT-hasA The exogenous gene can express Cry3AaT protein exogenously, and can achieve the synergistic effect of Yersinia insectivora CSLH88 and Cry3AaT insecticidal toxin protein, can be used in the prevention and control of coleopteran pests, and has significant insecticidal activity.

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

The invention belongs to the technical field of biochemistry, and particularly relates to a jellyfish punching toxin protein as well as a separation and purification method and application thereof. According to the method, a Superdex 75 16 / 600 GL gel filtration chromatographic column and a Mono Q anion exchange chromatographic column are used for separating the toxin protein from the sandfish venom for the first time, and the toxin protein can form pores in the surface of a cell membrane. The punching toxin protein obtained by the invention lays a foundation for deeply researching the structure and mechanism of the punching toxin protein of the sandfish and developing a medicine for treating the stings of the sandfish.

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.