39 results about "Cytoplasmic Polyhedrosis Viruses" patented technology

The invention relates to disinfectant for a rearing instrument of a rearing house and application of the disinfectant. The disinfectant for the rearing instrument of the rearing house comprises the following components in parts by weight: 1 part of chlorobromoisocyanaria acid, 2-10 parts of alkaline sodium salt and 500-2,000 parts of water, wherein the pH (Potential Of Hydrogen) is 9.0-12.0, and the disinfectant is prepared by uniformly mixing the material components. According to the invention, the defects that the chlorobromoisocyanaria acid applied to silkworm breeding disinfection prevention cannot kill bombyx mori nuclear polyhydrosis virus (BmNPV) and bombyx mori cytoplasmic polyhedrosis virus (BmCPV) are solved; and the product can kill various silkworm pathogenic bacteria after being applied once, and other drugs are not needed to apply.

The invention relates to the technology of antigen protein expression, and particularly relates to a method for preparing a cyprini herpesvirus II antigen coated polyhedrosis based on a baculovirus expression system. According to the method, by designing and recombining bombyx mori nuclear polyhydrosis virus BmNPV-VP3-cyHV-polh, 1-186, 993-1197, 603-783 and 85-186 regional sequences of ORF72, ORF66, ORF81 and ORF82 and the coding sequence of the 1-279 region of a VP3 gene of bombyx mori cytoplasmic polyhedrosis virus structural protein are connected in series to form a fused sequence which is controlled by baculovirus P10 promoter; and the bombyx mori cytoplasmic polyhedrosis protein gene is controlled by a polyhedrin gene promoter of baculovirus. The virus is used for inoculating bombyx mori or bombyx mori culture cells, the recombinant virus expressed cyprini herpesvirus II antigen protein can be coated in bombyx mori cytoplasmic polyhedrosis; the formed polyhedrosis can be purified by simple differential centrifugation; the purified polyhedrosis is cracked under a basic condition, the polyhedrosis protein can be precipitated by centrifuging, and the cyprini herpesvirus II antigen is reserved in supernatant, so that the cyprini herpesvirus II antigen can be quickly and conveniently obtained.

It is intended to provide a protein complex and a production process whereby the protein complex can be efficiently produced without lowering its function. It is also intended to provide use of the protein complex in a biosensor, an immobilized enzyme and so on. A protein complex comprising a polyhedral protein having an insect virus encapsulated therein and a target protein having a restricted region of a capsid-protein VP3 of cytoplasmic polyhedrosis virus, more specifically, a region which is either a region from the N-terminus to the 40th amino acid residue or a region from the 41st amino acid residue to the 79th amino acid residue as an embedding signal for polyhedron, and a process for producing the same. The polyhedral protein has an effect on improvement in the stability of the target protein, protection thereof or improvement in the preservation properties thereof, or a combination of any of these. The target protein is at least one member selected from the group consisting of fluorescent or light-emitting proteins, enzymes, antigens, antibodies, cytokines, receptors and bioactive proteins. A biosensor characterized in that the above-described protein complex is arranged in dots or lines on a substrate and immobilized thereon.

The invention discloses a Chinese cabbage pest and disease damage preventing agent. The Chinese cabbage pest and disease damage preventing agent is prepared from sophora flavescens, common cnidium fruit, snake gourd fruit, common anemarrhena rhizomes, garden burnet, Chinaberry seeds and leaves, tea seeds, strain fermentation liquor, green muscardine fungi, modified diatomite and cytoplasmic polyhedrosis viruses. The agent is mainly prepared from Chinese herbal medicine, prepared liquid medicine contains toosendanin, sophocarpidine, tea saponin and other substances and has a high inhibiting effect on various germs, the incidence rate of downy mildew, anthracnose and other diseases of Chinese cabbages is reduced, the modified diatomite can adhere to the epidermis or shells of pests, tissue is softened, the green muscardine fungi and the cytoplasmic polyhedrosis viruses are helped to invade and be parasitized into the bodies of pests, and the pest killing effect is enhanced. The components of the agent interact with one another, the incidence rate of pest and disease damage of Chinese cabbages is greatly reduced, the components are safe and free of pollution, the quality of crops is improved, and popularization value is high.

The invention discloses a primer and method for detecting a bombyx mori cytoplasmic polyhedrosis virus (BmCPV). The invention provides a pair of specific primers, i.e., BmCPVf and BmCPVr. A RT-PCR (Reverse Transcription-Polymerase Chain Reaction) amplification program is designed, so that accurate and quick detection of the bombyx mori cytoplasmic polyhedrosis virus is realized. The method has high specificity, is quick and accurate for detection, is suitable for a small amount of sample operation, and is easy to operate. By adopting the method, the requirement on the molecular diagnosis of the BmCPV can be met, and technical bases are provided for early detection of production and prediction of midgut polyhedrosis of bombyx mori.

The invention discloses a method for realizing efficient covering of a foreign protein by using an insect virus polyhedron, wherein the polyhedrin of a bombyx mori cytoplasmic polyhedrosis virus (BmCPV) and a target protein gene are constructed under the PH and P10 promoters on a pFastBac Dual carrier, and a baculovirus capable of expressing the polyhedron and the target protein simultaneously can be obtained through recombination. The method is characterized in that the cytoplasmic polyhedrosis protein gene and the target protein gene are simultaneously constructed under the PH and P10 promoters on the pFastBac TMDual carrier through a baculovirus expression system, respectively, a flexible link peptide is introduced between the target protein and a signal peptide, and the recombinant baculovirus is obtained through recombination with a virus skeleton. Once Sf9 or Sf21 insect cells are infected by the single virtus, the desired covered protein can be directly generated in cells; the recombinant virtus also can be amplified.

The invention discloses a pine moth cytoplasmic polyhedrosis virus and beauveria bassiana insecticide suspension as well as a preparation method and application thereof. The pine moth cytoplasmic polyhedrosis virus and beauveria bassiana insecticide suspension disclosed by the invention comprises effective active components, wherein the active components comprise a pine moth cytoplasmic polyhedrosis virus suspension and beauveria bassiana powder; the active components account for 2-30% of the total mass of the whole preparation. The suspension is safe and non-toxic, has the characteristics ofhigh efficiency and broad spectrum, is harmless to livestock and human beings and is harmless to natural enemies.

The invention discloses an in vitro construction method of a cytoplasmic polyhedrosis virus of Bombyx mori. Particularly, the in vitro construction method comprises the following steps: 1) obtaining a cytoplasmic polyhedrosis virus genome of the Bombyx mori; 2) designing and combining primers; 3) obtaining cDNA of S1-S10 fragments of the cytoplasmic polyhedrosis virus genome; 4) carrying out PCR amplification on the cDNA; 5) carrying out endonuclease digestion on an amplified product, and cloning into a plasmid vector to obtain a recombinant plasmid; 6) after carrying out endonuclease digestion and linearization on the recombinant plasmid, obtaining RNA by virtue of in vitro transcription; and 7) equal mol mixing RNA, wrapping by using a liposome, transfecting a host, and thus obtaining the cytoplasmic polyhedrosis virus of Bombyx mori constructed in vitro. The in vitro construction method of the cytoplasmic polyhedrosis virus of the Bombyx mori has the action of promoting function research of each fragment of the cytoplasmic polyhedrosis virus genome and can also provide technical support to develop a recombinant cytoplasmic polyhedrosis virus biological insecticide.

The invention discloses a method for constructing a recombinant bombyx mori cypovirus for expressing a red fluorescence protein. Particularly, the method comprises the following steps: (1) obtaining a viral genome; (2) designing and synthesizing a primer; (3) obtaining a cDNA of the virus genome; (4) performing PCR amplification on the cDNA; (5) performing enzyme digestion on the amplified product to obtain a recombinant plasmid; (6) performing enzyme digestion on the recombinant plasmid pT-S1 to pT-S9 and performing in vitro transcription to obtain the corresponding RNA; (7) constructing a recombinant plasmid pT-S10-DsRed with red fluorescence protein to obtain the S10-DsRed RNA; and (8) mixing the RNA with equal mol, packaging and transfecting a host to obtain the virus. By adopting the method constructed by the invention, the red fluorescence protein can be expressed in the silkworm cell so that the required recombinant cytoplasmic polyhedrosis virus biological insecticides can be obtained.

The invention discloses a pesticide suspending agent containing CPV (cytoplasmic polyhedrosis virus) and beauveria bassiana. The pesticide suspending agent is characterized by comprising effective active components Dendrolimus punctatus cytoplasmic polyhedrosis virus suspension and beauveria bassiana powder, and the weight ratio of the Dendrolimus punctatus cytoplasmic polyhedrosis virus suspension to the beauveria bassiana powder is (1:12)-(2:1). The invention further discloses a preparation method of the pesticide suspending agent. The pesticide suspending agent is efficient, broad-spectrum, harmless to people and livestock, harmless to natural enemies and widely applicable to the prevention and treatment of pests of forests, vegetables, cotton, tea leaves, fruit trees and the like.

To provide a useful objective protein coated with a protective protein, a protein complex that is produced in cells and has a structure that the objective protein is occluded with the viral occlusion body protein. A protein complex having the structure that the objective protein is occluded with the viral occlusion body protein, produced in insect cells or plant cells, preferably produced by incorporation of the objective protein in the crystalline form during crystallization of polyhedron derived from polyhedrosis virus, preferably a protein complex in which the polyhedrin contributes to improvement of stability, or protection, or improvement of preservability, or combination thereof, of the objective protein.

The invention discloses a fluorescence quantitative polymerase chain reaction (PCR) detecting method for a silkworm cytoplasmic polyhedrosis virus, which belongs to the technical field of biology. The method includes construction of standard plasmids, design of specificity primers, amplification of real-time fluorescence quantitive PCR and building of a standard curve, extraction of ribonucleic acid (RNA) of a sample infected by the virus, preparation of complementary deoxyribonucleic acid (cDNA) and judging of detecting results. The fluorescence quantitative PCR detecting method is high in flexibility, good in stability, strong in specificity, capable of being quantitative and suitable for mass detecting of virus infection.

The invention relates to a detection primer and a detection method for detecting bombyx mori cytoplasmic polyhedrosis virus (BmCPV). Nucleotide sequences of the detection primer are as shown in SEQ ID NO.1 and SEQ ID NO.2. By means of the specific primer, a specific segment from the 366th base to the 736th base of RNA-dependent RNA polymerase (RDRP) gene from 5' to 3' of the BmCPV can be effectively amplified, the size of the segment is 370bp, and the specific primer is used for detecting the RDRP gene of the BmCPV, detection sensitivity is high, operation is simple, and by means of the detection primer and the detection method for detecting the BmCPV, important technical foundation is provided for control effect of silkworm diseases caused by infection of the BmCPV.

The invention belongs to the technical field of disease and pest control and particularly relates to a method for controlling dendrolimus punctatus by using artificial migration natural enemies. The method is characterized in that the method is based on a dendrolimus punctatus CPV (cytoplasmic polyhedrosis virus) oil stock solution, the dendrolimus punctatus CPV oil stock solution and a virus preparation prepared from honey are prepared and placed in a chigger cage containing dendrolimus punctatus cocoons, and thus the dendrolimus punctatus disaster is controlled. The method for controlling dendrolimus punctatus by using the migration enemies in combination with the dendrolimus punctatus virus has the technical advantages of quick and good dendrolimus punctatus control effect.

The invention discloses a bombyx mori cytoplasmic polyhedrosis virus RT-LAMP detection primer group and a detection kit thereof. The kit comprises the detection primer group, a BstDNA polymerase, a reaction solution, a stabilization solution, a coloring solution and a positive contrast solution, the detection primer group comprises CPVF2, CPVB2, CPVFI2 and CPVBI2 primers, and the nucleotide sequences of the CPVF2, CPVB2, CPVFI2 and CPVBI2 primers are represented by SEQIDNO:1-4 respectively. The accurate and rapid detection of the bombyx mori cytoplasmic polyhedrosis virus can be realized based on the kit. The kit has the advantages of high detection sensitivity, strong specificity, rapid and accurate detection, suitableness for the operation of a small amount of samples, and simple operation, provides a technical basis for the early-stage detection and prevision inspection of the bombyx mori midgut polyhedrosis in the production, and is of great practical significance.

The invention discloses a dendrolimus punctatus cytoplasmic polyhedrosis virus oiling agent, and a preparation method and application thereof. The dendrolimus punctatus cytoplasmic polyhedrosis virus oiling agent is prepared by soaking, chopping, eluting, filtering, centrifugally separating, drying in vacuum, blending and sub-packaging. A dendrolimus punctatus cytoplasmic polyhedrosis virus concentrated liquid is obtained by vacuum drying at 28-35 DEG C in negative 0.9 barometric pressure for 10-15 hours by virtue of a vacuum dryer; and the foul odor of the oiling agent is removed, so that a product is easy to popularize and apply; blending comprises the step of adding saponated glycerinum and analytically pure activated carbon; the content of a dendrolimus punctatus cytoplasmic polyhedrosis virus in the prepared oiling agent is 1500,000,000PIB / ml or above; the pathogenicity of the virus is high; and the expiration date of the oiling agent is prolonged and can reach 3-4 years. By virtue of the oiling agent disclosed by the invention, the infective rate of the dendrolimus punctatus cytoplasmic polyhedrosis virus can reach 95% or above; the oiling agent has a lasting control effect on pine moths in a forest for multiple years after prevention and treatment is performed once; and the lasting control effect can last for five years or above.

The invention relates to the biotechnology field and the technical field of biological medicines, and particularly relates to an expression method of an antibacterial peptide Hclocidin18 in escherichia coli. The expression method is characterized by comprising following steps of: a. plasmid construction, wherein the escherichia coli is utilized to perform recombinant plasmid construction, and a polyhedron of a bombyx mori cytoplasmic polyhedrosis virus (BmCPV) is utilized as a fusion partner; b. induction expression; c. purification of a fusion protein coded by the recombinant plasmid; and d. schizolysis of the fusion protein and collection of the antibacterial peptide Hclocidin18. The polyhedron of the bombyx mori cytoplasmic polyhedrosis virus (BmCPV) is utilized as the fusion partner to achieve success expression of the antibacterial peptide Hclocidin18 in the escherichia coli. The alkali dissolution characteristic of the polyhedron is utilized to achieve efficient recovery of the antibacterial peptide Hclocidin18.

A kind of production method of pine caterpillar type polyhedrosis virus oil preparation, the viral polyhedron of susceptible pine caterpillar midgut is separated from worm body, and separates from other solid phase substances, then process and concentrate, add solvent ethylene glycol, The suspending agent sodium dodecylbenzene sulfonate is prepared by mixing, stirring and grinding. Its pine caterpillar polyhedrosis virus has high purity, small particle size, and good suspension effect, which solves the problems of sedimentation and layering and blocking nozzles, thereby meeting the requirements of ultra-low volume spraying. The pine caterpillar polyhedrosis virus made according to this method Body virus oil agent, insecticidal rate of more than 92%, its production cost is low, does not pollute the environment, is safe for humans and animals, and has good economic and social benefits.

The invention relates to the field of immobilized protein, in particular to a method for preparing heterologous protein wrapped polyhedrosis. The bombyx mori cytoplasmic polyhedrosis protein, a fusion foreign protein with a bombyx mori cytoplasmic polyhedrosis virus structural protein VP5 tag and a protease specific cleavage site are co-expressed in bombyx mori culture cells or bombyx mori by using a baculovirus expression system through a genetic engineering method, so that the polyhedrosis wrapping the foreign protein is formed. The formed polyhedron can be purified through simple differential centrifugation; the polyhedrin has protection and slow release effects on foreign proteins wrapped by the polyhedrin. After the purified polyhedrosis is cracked under an alkaline condition, the pH value is adjusted to a polyhedrosis protein isoelectric point by using a hydrochloric acid solution, the polyhedrosis protein can be precipitated through centrifugation, and the fusion foreign protein is retained in a supernatant, so that the fusion foreign protein can be quickly and conveniently obtained; the fusion foreign protein is subjected to enzyme digestion through corresponding protease to remove a VP5 tag, so that a foreign protein closer to the nature can be conveniently obtained.

The invention relates to the technology of antigen protein expression, and particularly relates to a method for preparing a cyprini herpesvirus II antigen coated polyhedrosis based on a baculovirus expression system. According to the method, by designing and recombining bombyx mori nuclear polyhydrosis virus BmNPV-VP3-cyHV-polh, 1-186, 993-1197, 603-783 and 85-186 regional sequences of ORF72, ORF66, ORF81 and ORF82 and the coding sequence of the 1-279 region of a VP3 gene of bombyx mori cytoplasmic polyhedrosis virus structural protein are connected in series to form a fused sequence which is controlled by baculovirus P10 promoter; and the bombyx mori cytoplasmic polyhedrosis protein gene is controlled by a polyhedrin gene promoter of baculovirus. The virus is used for inoculating bombyx mori or bombyx mori culture cells, the recombinant virus expressed cyprini herpesvirus II antigen protein can be coated in bombyx mori cytoplasmic polyhedrosis; the formed polyhedrosis can be purified by simple differential centrifugation; the purified polyhedrosis is cracked under a basic condition, the polyhedrosis protein can be precipitated by centrifuging, and the cyprini herpesvirus II antigen is reserved in supernatant, so that the cyprini herpesvirus II antigen can be quickly and conveniently obtained.

The invention discloses a clanis bilineata tsingtauica cytoplasmic polyhedrosis virus strain and a proliferation method thereof. The virus strain is clanis bilineata tsingtauica cytoplasmic polyhedrosis virus, and the preservation name is clanis bilineata tsingtauica cytoplasmic polyhedrosis virus CbCPV. The clanis bilineata tsingtauica cytoplasmic polyhedrosis virus can effectively control clanis bilineata tsingtauica harm, continuously acts after one-time application, can maintain no clanis bilineata tsingtauica harm for two to three years, is convenient to produce, reduces the use of chemical pesticides, is environment-friendly, and can greatly reduce the clanis bilineata tsingtauica harm in soybean production, which assists the work of grain battling with insects in soybean production in China.

The invention relates to disinfectant for a rearing instrument of a rearing house and application of the disinfectant. The disinfectant for the rearing instrument of the rearing house comprises the following components in parts by weight: 1 part of chlorobromoisocyanaria acid, 2-10 parts of alkaline sodium salt and 500-2,000 parts of water, wherein the pH (Potential Of Hydrogen) is 9.0-12.0, and the disinfectant is prepared by uniformly mixing the material components. According to the invention, the defects that the chlorobromoisocyanaria acid applied to silkworm breeding disinfection prevention cannot kill bombyx mori nuclear polyhydrosis virus (BmNPV) and bombyx mori cytoplasmic polyhedrosis virus (BmCPV) are solved; and the product can kill various silkworm pathogenic bacteria after being applied once, and other drugs are not needed to apply.

The invention relates to the field of pesticides, and particularly relates to diatomite-pine moth cytoplasmic polyhedrosis virus wettable powder. The wettable powder at least comprises a pharmaceutical composition, a wetting agent, a dispersing agent and filler, wherein the pharmaceutical composition comprises diatomite and pine moth cytoplasmic polyhedrosis virus in a weight ratio of (1: 60)-(400: 1), the microstructure of the diatomite is of a sawtooth structure, the dispersing agent is alkyl naphthalene sulfonate formaldehyde condensate and glucose in a mass ratio of (1-3): 1, and the filler is preferably sodium alkyl naphthalene sulfonate. The diatomite-pine moth cytoplasmic polyhedrosis virus wettable powder provided by the invention solves the problems that the pine moth cytoplasmicpolyhedrosis virus takes effect slowly, the diatomite does not have surface activity and the like, and the powder is good in heat storage stability, uniform, loose, not easy to agglomerate and cake, easy to disperse and wet after being diluted with water, high in suspension rate and relatively good in infiltration spreadability on plant surfaces.

The invention discloses an in vitro construction method of silkworm plasmopolyhedrosis virus. Specifically, the method of the present invention comprises the following steps: 1) Obtaining the genome of Bombyx mori plasmopolyhedrosis virus; 2) Designing and synthesizing primers; 3) Obtaining the cDNA of fragments S1 to S10 of the viral genome; 4) Performing PCR amplification on the cDNA ;5) Digest the amplified product, and then clone it into a plasmid vector to obtain a recombinant plasmid; 6) Digest and linearize the recombinant plasmid, and transcribe in vitro to obtain RNA; and 7) Mix the RNA equimolarly , use liposome to encapsulate and transfect the host, and then obtain the silkworm plasmopolyhedrosis virus constructed in vitro. The in vitro construction method of the silkworm polyhedrosis virus of the present invention not only promotes the functional research of each fragment of the silkworm polyhedrosis virus genome, but also provides technical support for the research and development of the recombinant cytoplasmic polyhedrosis virus biopesticide.

The invention discloses an insecticidal composition containing fluchlorabendiamide and a bio-sourced insecticide. Effective active components comprise an active component A and an active component B,wherein the active component A is the fluchlorabendiamide; the active component B is selected from any one of abamectin, spinosad, spinetoram, bacillus thuringiensis, milbemectin, emamectin, emamectinbenzoate, sucrose octacarboxylate, beauveria bassiana, metarhizium anisopliae, nuclear polyhedrosis virus, cytoplasmic polyhedrosis virus and granulosis virus; wherein the weight ratio of the activecomponent A to the active component B is 1:80-80:1. The composition disclosed by the invention has a synergistic effect on various pests harmful to agricultural production, the dosage of pesticides isreduced, the residual quantity of the pesticides on crops is reduced, the environmental pollution is reduced, the composition is safe to human and livestock and good in environmental compatibility, and the generation of pest resistance is delayed.

The invention discloses a DNA carrier based in vitro construction method of a nombyx mori cytoplasmic polyhedrosis virus. Specifically, the method comprises the following steps: 1) acquiring a genomeof a nombyx mori cytoplasmic polyhedrosis virus; 2) designing and synthesizing a primer; 3) acquiring cDNA of fragments S1 to S10 of the genome of the virus; 4) conducting PCR (Polymerase Chain Reaction) amplification on the cDNA; 5) implementing enzyme digestion on an amplification product, then cloning to a plasmid vector, acquiring a recombinant plasmid; 6) implementing enzyme digestion and linearization on the recombinant plasmid; 7) mixing the linearized plasmid at equal molar ratio, wrapping by using a lipidosome, transfecting and expressing a host cell of a T7-RNA polymerase, and further acquiring the nombyx mori cytoplasmic polyhedrosis virus constructed in vitro. The DNA carrier based in vitro construction method of the nombyx mori cytoplasmic polyhedrosis virus not only has an effect on promoting the functional study of various fragments of the genome of the nombyx mori cytoplasmic polyhedrosis virus, but also provides a novel method for acquiring the nombyx mori cytoplasmicpolyhedrosis virus in vitro, and provides theoretical direction and technical support for the researches and development of the construction of a recombinant cytoplasmic polyhedrosis virus biopesticide.

The invention discloses a fluorescence quantitative polymerase chain reaction (PCR) detecting method for a silkworm cytoplasmic polyhedrosis virus, which belongs to the technical field of biology. The method includes construction of standard plasmids, design of specificity primers, amplification of real-time fluorescence quantitive PCR and building of a standard curve, extraction of ribonucleic acid (RNA) of a sample infected by the virus, preparation of complementary deoxyribonucleic acid (cDNA) and judging of detecting results. The fluorescence quantitative PCR detecting method is high in flexibility, good in stability, strong in specificity, capable of being quantitative and suitable for mass detecting of virus infection.