116 results about "Toxic proteins" patented technology

Disclosed are Bacillus thuringiensis strains comprising novel crystal proteins which exhibit insecticidal activity against coleopteran insects including red flour beetle larvae (Tribolium castaneum) and Japanese beetle larvae (Popillia japonica). Also disclosed are novel B. thuringiensis crystal toxin genes, designated cryET33 and cryET34, which encode respectively the coleopteran-toxic proteins, CryET33 (29-kDa) crystal protein, and CryET34 (14-kDa) crystal protein. Also disclosed are methods of making and using transgenic cells comprising the novel nucleic acid sequences of the invention.

The invention provides compositions comprising polynucleotide molecules encoding certain pesticidal polypeptides which exhibit plant parasitic nematode and / or insect control properties, and are particularly directed to controlling plant parasitic pest species of nematodes and insects known to infest crop plant species. Methods for controlling pests are disclosed in which the toxic proteins are provided in the diet of the targeted plant pests. The invention also provides compositions such as nucleic acids, proteins, and plant and bacterial cells, plants, and seeds containing the nucleic acid and protein compositions, as well as methods and kits for identifying, detecting, and isolating the compositions of the present invention. The invention further provides a method of producing crops from recombinant seeds which contain the polynucleotide molecules encoding the pesticidal polypeptides of the present invention.

The invention provides a method of propagating an adenoviral vector. The method comprises (a) providing a cell comprising a cellular genome comprising a nucleic acid sequence encoding a tetracycline operon repressor protein (tetR), and (b) contacting the cell with an adenoviral vector comprising a heterologous nucleic acid sequence encoding a toxic protein. The heterologous nucleic acid sequence is operably linked to a promoter and one or more tetracycline operon operator sequences (tetO), and expression of the heterologous nucleic acid sequence is inhibited in the presence of tetR, such that the adenoviral vector is propagated. The invention also provides a system comprising the aforementioned cell and adenoviral vector.

A self-cleaving element for use in bioseparations has been derived from a naturally occurring, 43 kDa protein splicing element (intein) through a combination of protein engineering and random mutagenesis. A mini-intein (18 kDa) previously engineered for reduced size had compromised activity and was therefore subjected to random mutagenesis and genetic selection. In one selection a mini-intein was isolated with restored splicing activity, while in another, a mutant was isolated with enhanced, pH-sensitive C-terminal cleavage activity. The enhanced cleavage mutant has utility in affinity fusion-based protein purification. The enhanced splicing mutant has utility in purification of proteins such as toxic proteins, for example, by inactivation with the intein in a specific region and controllable splicing. These mutants also provide new insights into the structural and functional roles of some conserved residues in protein splicing. Thus, disclosed and claimed are: a genetic system and self-cleaving inteins therefrom; bioseparations employing same; protein purification by inactivation with inteins in specific regions and controllable intein splicing; methods for determining critical, generalizable residues for varying intein activity; and products.

The invention provides an insect-resistant gene Cry1Ab-Material. The synthesis of the gene is as follows: according to the amino acid sequence of the N-terminal of the original Bacillus thuringiensis (Bt) protein (Cry1Ab), the preferred codons of monocotyledon (corn) is used to perform human reformation and synthetize a new Cry1Ab DNA sequence; and the prokaryotic expression vector and plant expression vector are constructed, and transformation is performed in the host cells. Vitro tests prove that the transformed and synthetized Bt gene toxic protein has obvious insecticidal effect on Ostrinia nubilalis. The expression of the insect-resistant gene Cry1Ab-Ma in monocotyledon is stable and efficient, thus the gene can be used to produce insect-resistant transgenic plant.

This invention relates to a method for increasing the conversion efficiency of pollen tube pathway method by using agrobacterium tumefaciens virulence gene. The method comprises: combining or mixing agrobacterium tumefaciens virulence gene and the target gene to be converted (green fluorescent protein gene) in vitro, packaging with liposome, and converting plants, especially cotton by using pollen tube pathway method. The method combines agrobacterium-liposome-pollen tube pathway technique, and has obviously increased protoplast conversion efficiency. Plasmids are not only protected by liposome when entering embryo sacs, but also encapsulated by the toxic protein to be expressed when entering egg cells, which can improve the stability of exogenous DAN in cell conversion process, realize precise cutting in T-DNA region, and increase the conversion efficiency.

The invention discloses drug-resistant coli phage and application thereof. The classification of the drug-resistant coli phage is named as Escherichia phage vB_EcoP-EG1, the reservation number is CCTCC M 2018562, and the preservation date is August 21, 2018. The phage Escherichia phage vB_EcoP-EG1 has stable properties, a short life cycle and high lysis ability to host bacteria. The results of a genome and a protein map show that the phage Escherichia phage vB_EcoP-EG1 does not encode toxic proteins. At the same time, the phage Escherichia phage vB_EcoP-EG1 can infect and lyse a plurality of strains of clinically and randomly isolated multi-drug resistant escherichia coli, and particularly has a good killing effect on uropathogenic escherichia coli. In conclusion, Escherichia phage vB_EcoP-EG1 is a good candidate phage for phage therapy, can be used as a single preparation or mixed with other phages to form a cocktail preparation for fighting against infection caused by the clinical multi-drug resistant escherichia coli.

Methods and compositions are provided for producing membrane proteins or toxic proteins from recombinant DNA introduced into a prokaryotic host cell by targeting the expressed proteins to the envelope of the host cell. The methods and compositions utilize a protein vehicle fused to a protein of interest. The fusion protein may contain one or more protease cleavage sites to separate the protein of interest from the protein vehicle either in vivo or in vitro. The protein vehicle is characterized by a membrane-tar peptide and a trans-membrane segment separated by a cytoplasmic amino acid sequence that includes a cytoplasmic affinity-binding domain.

The present invention relates to an expression vector comprising, in 5′ to 3′ direction, a promoter, a multiple cloning site and a nucleotide sequence encoding glutathione-S-transferase (GST), for production of a fusion protein comprising a membrane protein, secretory protein or toxic protein or peptide, fused directly or indirectly with the N-terminal of GST. Preferably, the fusion protein comprises GST and a membrane protein or membrane localised peptide. The invention is especially suitable for membrane proteins having their C-terminals in the cytoplasm. The invention also relates to methods for producing such fusion proteins using host cells transformed with the expression vector in which a desired gene has been cloned.

This invention relates to a method of controlling pathogens, by use of oxalate-producing enzyme, alone or in combination with toxic protein may be applied directly to the plant or produced thereon by microorganisms or by genetically modifying the plant to produce the enzyme, and to genes, microorganisms, and plants useful in that method. In one aspect this invention relates to a method of controlling insects, including lepidopterans and boll weevils, by use of oxalate-producing enzyme, alone or in combination with a crystal protein from Bacillus thuringiensis or cholesterol oxidase which may be applied directly to the plant or produced thereon by microorganisms or by genetically modifying the plant to produce the enzyme, and to genes, microorganisms, and plants useful in that method.

The invention discloses a maize breeding method. A BT insecticidal toxic protein gene, a bar gene and maize dwarf mosaic virus coat protein gene MDMVCPDNA are introduced into embryogenic maize calluses through particle bombardment, so that seeds with excellent insect resistance, weed resistance, disease resistance and drought tolerance are obtained; through seed soaking treatment by seed-soaking liquid, germination of seeds is promoted, the respiration rate of seedlings is increased, the root activity of the seedlings is improved, and the resistance of both the seeds and the seedlings is enhanced; through drip irrigation of a drought-resistant nutrient and by using anti-oxidation substances of phenolic acids, the number and activity of the protective enzymes in the plants are improved, the content of polyphenol substances is increased, the content of anti-oxidation substances in the crop is increased and the drought resistance of the crop is enhanced; meanwhile, since the content of the anti-oxidation substances in the crop is increased, the disease resistance, UV radiation resistance and the like of the crop are improved; since monopotassium phosphate and nitrogen are added into the phenol substances, nutrient elements supplied to the crop are increased.

The present invention provides an improved process for the production of recombinant peptides by fusion of recombinant peptides with icosahedral viral capsids and expression of the fusion in bacterial cells of Pseudomonad origin. The Pseudomonad cells support formation of virus like particles from icosahedral viral capsids in vivo, and allow the inclusion of larger recombinant peptides as monomers or concatamers in the virus like particle. The invention specifically provides cells expressing viral capsid fusions, nucleic acids encoding fusions of toxic proteins with icosahedral viral capsids and processes for manufacture of recombinant proteins.

The invention relates to a marker protein expression cassette capable of inducing regulation and a recombinant vector constructed by the same and an application. A selection marker protein expression cassette capable of inducing regulation is constructed by use of selection marker protein and a regulation induction element; a recombinant vector taking escherichia coli toxic protein as a selection marker is prepared through the selection marker protein expression cassette capable of inducing regulation; the recombinant vector controls to express escherichia coli toxic protein in secondary homologous recombination; the strain only experiencing once homologous recombination is killed or the growth is inhibited while the growth of the strain experiencing twice homologous recombination is not limited, so that the selection efficiency of the strain of twice homologous recombination is improved, the operation flow of gene knock-in or knockout of a bacillus homologous recombination method is simplified, the operation difficulty is lowered, the labor intensity is reduced, and the marker protein expression cassette can be perfectly used for knocking out the bacillus gene and transferring into the bacillus together with exogenous gene.

The present invention relates to a system for expressing toxic proteins, to an expression vector comprising this system, to a prokaryotic cell transformed with this system, and also to a method for synthesizing a toxic protein using this expression system. The expression system of the invention is characterized in that it comprises successively, in the 5′-3′ direction, a nucleotide sequence encoding the Asp-Pro dipeptide and a nucleotide sequence encoding a toxic protein. According to a preferred embodiment of the invention, the expression system also comprises, upstream of the Asp-Pro sequence, a nucleotide sequence encoding a soluble protein. The expression system of the invention makes it possible to construct an expression vector that is useful for transforming a prokaryotic cell such as E. coli, for example in a method for synthesizing the toxic protein.

The present invention relates to a composition comprising an autophagy enhancement compound. Small molecules that are able to enhance autophagy and lysosome biogenesis by activating the gene TFEB which can prevent the accumulation of toxic protein aggregates in treating neurodegenerative diseases are disclosed.

The invention relates to specifically relates to a plasmid vector capable of realizing directional TA cloning, background sticky end-free cloning and expression, and a construction method thereof, belonging to the field of gene engineering. The plasmid vector is a closed circular double-stranded DNA and contains two multiple cloning sites, wherein a ccdB expression cassette exist between the two multiple cloning sites, and the ccdB expression cassette can realize constitutive expression of toxic protein ccdB, which is used as a negative selection marker, in most Escherichia coli strains. The multiple cloning sites are designed on the basis of the plasmid vector; and the plasmid vector is applicable to directional TA cloning and improved background sticky end-free cloning. Moreover, the plasmid vector is directly applicable to expression and purification of proteins. Each function of the plasmid vector is verified. The plasmid vector provided by the invention is expected to play an important role in the field of gene engineering due to its convenience and high efficiency.

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.

A process for extracting the scorpion venom and cleaning it includes such steps as cleaning the living scorpion, stimulating it at a temp higher than 30 deg.C in aseptic environment by electric pulses (8-10 v) at 139 Hz for a time less than 1 s, collecting scorpion venom by a container at -2-2 deg.C, filtering and freeze-drying. Its advantages are high quality and content of toxic protein, and low damage to scorpion.

The invention provides a citrus canker susceptible gene CsLOB. The sequence of the citrus canker susceptible gene CsLOB in citrus is as shown in SEQ ID No.1, the sequence length of a promoter is 500bp, and the sequence length of an ORF (Open Reading Frame) is 711bp; and the sequence of CsLOB protein is as shown in SEQ ID No.2. UPT box sequence which can be specifically combined with a citrus canker bacterium PthA exists in the CsLOB gene promoter, the sequence of the UPT box is as shown in SEQ ID No.3, and the length of the UPT box is 18pb. The obtained CsLOB gene can be specifically recognized by main toxic protein PthA derived from the citrus canker bacterium and is activated to be transcribed, so that the occurrence of citrus canker is caused. A new germplasm of citrus or a new material with durable resistance can be obtained through gene silencing of the CsLOB gene and genetic modification of the UPT box.

The invention discloses a method of determining bacillus thuringiensis toxic protein CrylAc. The method comprises the steps that: Fe3O4@Au nana-particles combining with a transgenic Bt toxic protein CrylAc primary antibody is modified onto the surface of a magnetic control glassy carbon electrode, and specific immune reaction is carried out on the magnetic control glassy carbon electrode with an antigen and a GOD labeled secondary antibody to form a compound with a sandwich structure; an electrogenerated chemiluminescence immunosensor is developed to be used for detecting the concentration of transgenic Bt toxic protein CrylAc; glucose is catalyzed and oxidized by using GOD to generate H2O2; voltage is applied to a working electrode to excite luminal to be oxidized; a luminal oxide and the H2O2 react to generate an electrochemiluminescence signal; an electrochemiluminescence response is strengthened along with the increase of the concentration of the GOD; and the electrochemiluminescence signal has a linear relation with the concentration of the transgenic Bt toxic protein CrylAc within a range of 0-6ng / mL. The method has high sensitivity and low detection limit, is widely applied and supplies a wide application prospect of reliably and ultra-sensitively detecting the toxic protein of a transgenic plant.

A method for testing trace Bt toxic protein in the bodies of insects in a transgenic Bt crop habitat includes the following steps: in the first step, collecting insects to be tested growing in a transgenic Bt crop field, and preparing insect liquid to be tested; in the second step, collecting insects growing on non-transgenic Bt plants in a lab, which belong to the same species or variety as the insects to be tested, and preparing contrast insect liquid; in the third step, using the contrast insect liquid to prepare standard Bt toxic proteins with a series of concentrations; in the fourth step, utilizing an enzyme immunoassay method to determine the optical density values of the standard Bt toxic proteins and the insect liquid to be tested at the wavelength of 650 nanometers; in the fifth step, obtaining a standard equation according to the concentrations of the standard Bt toxic proteins and the corresponding optical density values, and calculating the Bt toxic protein concentration in the insect liquid to be tested according to the standard equation. The test method has extremely high sensitivity and good repeatability.

The invention discloses application of mercuride in preparing antivenom medicaments, aiming at solving the problems of unobvious treatment effect, high death rate and the like in the prior art, wherein the mercuride is one of mercuric sulfide, mercuric oxide, mercuric sulfate, mercuric chloride and the like. Because the main component in the mercuride is heavy metal which has the characteristic of inactivating all proteins, the mercuride has fatal destructive effect on all the proteins; and snake venom is formed by mixing dozens of poisonous proteins, therefore, the mercuride can inactivate the poisonous proteins in the snake venom so as to enable the poisonous proteins to lose activity (toxicity), thereby achieving the purpose of resisting the snake venom.

The present invention relates to bacterial expression vectors. In particular, the present invention provides tightly-regulated bacterial expression vectors designed for the cloning and expression of toxic proteins, RNA, and metabolites in vivo. The present invention thus provides methods of expressing protein and RNAs that were previously not able to be expressed.

The invention relates to an application of a compound 3,4,8,9,10-pentahydroxyl dibenzotriazole[b,d]pyran-6-actone in preparing an antibacterial drug. The compound is firstly separated from mature fruits of Burma terminalia citrina (Gaertin) Roxb. According to the experiment, the compound has relatively good inhibition effect on secretion of toxic proteins SipA, B, C and D of salmonella T3SS, doesnot have inhibition effect on growth of salmonella, and inhibits invasion, on HeLa cells, of salmonella by inhibiting transfer of toxic proteins. Therefore, the compound 3,4,8,9,10-pentahydroxyl dibenzotriazole[b,d]pyran-6-actone can be used for preparing the antibacterial drug, especially can be used for preparing a gram-negative pathogenic drug.

The invention relates to a method for inducing phytophthora capsici to generate toxic secretory proteins. The method comprises the following steps: bringing pepper phytophthora blight tissues back to a laboratory, scraping off the basal part surfaces of stems of diseased plants by a surgical knife blade at first, then washing the diseased plants by running water, taking 0.5cm of tissue blocks from disease-health junctions and placing the tissue blocks in a V8 culture medium, culturing for 3-4 days under a 25 DEG C dark condition until a large bacterial colony is formed, transferring a bacterial disk into a toxic protein liquid induction culture medium, carrying out shaking culture for 15-20 days, filtering mycelia by filter paper, centrifuging the filtrate for 30min at a speed of 8000rp, adding saturated ammonium sulphate in the supernatant, centrifuging the filtrate for 30min at a speed of 12000rp, carrying out a dialysis treatment on a precipitate, freeze-drying to obtain the toxic proteins secreted by the phytophthora capsici, and storing the toxic proteins at -70 DEG C. The method is capable of inducing the phytophthora capsici to generate lots of the toxic secretory proteins, thus filling in the gap that lots of toxic secretory proteins cannot be obtained in the domestic and overseas phytophthora capsici pathogenesis researches, and having great significance.

The invention discloses a method for preparing inositol by a multi-enzyme reaction system expressed by edible microorganisms. The method has the advantages that strains for the food industry serve asmulti-enzyme expression systems to produce isoamylase, glucan phosphorylase, glucose phosphate mutase, inositol-3-phosphate synthase and inositol monophosphatase, which are required for catalyzing starch and derivatives thereof, so that the possibility of contaminating the inositol by toxic protein, antigenic protein or endotoxin generated by escherichia coli in a production process is avoided fundamentally, a strict and complicated purification process is avoided, and the production cost is reduced.

The invention discloses a method for improving the transgenosis efficiency of cotton. The method comprises the following steps: after mixing restriction enzyme digestion DNA with a plasmid DNA protective agent, introducing a target gene into cotton through the pollen tube pathway method by using lipidosome packaging to carry toxic protein genes on Ti plasmids. The method can effectively improve the transgenosis efficiency.

The invention relates to the field of genetic engineering, and in particular to a plasmid vector, which is capable of flexibly implementing non-background TA cloning, cohesive end cloning and blunt end cloning and is capable of implementing temperature-induced protein expression, and a construction method of the plasmid vector. The plasmid vector is closed-ring shaped double-stranded DNA, which contains a gene, which is capable of coding toxic protein ccdB, as a negative selection marker. A flexible cloning function and a temperature-induced expression function of the plasmid are verified. Theplasmid, which is more convenient and efficient, can play an important role in the field of genetic engineering.