24 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 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.

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.

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.

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

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 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.

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 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 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.

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 provides a recombinant pertussis toxin S1 protein as well as a preparation method and application thereof. The amino acid sequence of the recombinant pertussis toxin S1 protein is as follows: (1) the amino acid sequence is as shown in SEQ ID NO.1; (2) is an amino acid sequence obtained by deletion, substitution or insertion of 1, 2, 3, 4 or more amino acids of the amino acid sequence as shown in SEQ ID NO.1; or (3) an amino acid sequence which has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher homology with the amino acid sequence as shown in SEQ ID NO.1. An antibody generated by induction of the recombinant pertussis toxin S1 protein as an immunogen is used as a detection primary antibody of a double-antibody sandwich method, the content of the complete pertussis toxin antigen can be accurately detected, and the detection method is more sensitive and efficient.

The invention discloses an engineered PVC protein compound for targeted removal of intracellular staphylococcus aureus as well as a preparation method and application of the engineered PVC protein compound, and belongs to the fields of genetic engineering, microbiology and drug delivery. Genetic engineering modification is carried out on PVC, a natural recognition domain of tail fiber protein of the PVC is replaced by a staphylococcus aureus receptor binding structural domain (SRapid BR), and meanwhile, natural toxin protein loaded in an inner cavity of the PVC is replaced by human beta-defensin-3 (HBD-3) with efficient bactericidal activity. The engineered compound can directly deliver HBD-3 into infected cells, effectively remove intracellular staphylococcus aureus which is difficult to kill by conventional antibiotics, and relieve inflammatory response caused by infection. The invention provides a brand-new treatment strategy for resisting intracellular bacterial infection with high targeting, and shows a wide application prospect as a programmable protein delivery platform.

An isolated polynucleotide which encodes a protein comprising a polypeptide sequence exhibiting at least 88% sequence identity to the amino acid sequence as set forth in SEQ ID NO.6 over a length of at least 300 amino acids and that can be used to inhibit Hemipteran insects.

Provided is an antibiotic resistance gene-free plasmid-containing production strain. The production strain is a gene-edited strain of the PIR strain, designated as PIR1-WN::0636 or PIR1-PR::0636. The production strain contains a nucleotide sequence encoding a toxin protein and an antibiotic resistance gene-free plasmid, wherein the antibiotic resistance-free plasmid contains a nucleotide sequence encoding an antitoxin protein; preferably, the replicon DNA element for the antibiotic resistance gene-free plasmid is R6K-γ. The toxin protein gene of the provided production strain can be stably maintained during strain passage, and has lethality upon induction, which can be used for plasmid screening. When the provided antibiotic resistance gene-free plasmid is transformed into the PIR1-WN::0636 strain, the positive rate reaches 80% or higher. Moreover, the antibiotic resistance gene-free plasmid can enable the stable production of plasmids with a high supercoiled proportion.