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182results about How to "Non-pathogenic" patented technology

Recombinant low-virulent vaccine strain of chicken infectious bursal disease viruses (IBDV) and application thereof

The invention discloses a recombinant low-virulent vaccine strain of chicken infectious bursal disease viruses (IBDV) and application thereof. In the invention, a major protective antigen gene VP2 of an epidemic superhigh virulent strain is cloned, the nucleotide of the gene VP2 is modified by mutation and then used for replacing a corresponding segment of a Gt genome of a low-virulent strain of the IBDV, so that the infectious clone of a recombinant genome of the IBDV is constructed, and the recombinant low-virulent vaccine strain is saved and identified by using an IBDV reverse genetic operation system. The microbial collection number of the vaccine strain is CGMCC No.3749. The recombinant low-virulent vaccine strain of the invention has high replicability, genetic stability and safety. The immune effect of the low-virulent vaccine strain of the invention is as good as that of the medium-virulent vaccine strain, but is superior to that of the low-virulent vaccine strain. The biological safety of the low-virulent vaccine strain of the invention is superior to that of the medium-virulent vaccine strain. As the vaccine strain, the recombinant low-virulent vaccine strain of the invention has the characteristics of high efficiency and low toxicity, is a good candidate vaccine strain and can be used for controlling chicken infectious bursal disease.
Owner:HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI

Grass carp reovirus S11 gene eukaryotic expression recombinant plasmid preparation method and application thereof in serving as nucleic acid vaccine

The invention discloses a grass carp reovirus S11 gene eukaryotic expression recombinant plasmid preparation method and application thereof in serving as nucleic acid vaccine, and belongs to the technical field of gene engineering and molecular immunology. The preparation method disclosed by the invention is characterized by comprising the steps of extracting viral genome RNA, performing reverse transcription to convert the viral genome RNA into cDNA, amplifying a corresponding DNA sequence out, constructing the DNA sequence to pcDNA-3.1(+) plasmid, converting Escherichia coli DH5alpha, screening out positive clone bacteria containing recombinant plasmid, culturing a lot of the positive bacteria and extracting recombinant plasmid S11-pcDNA3.1 contained in the bacteria. The recombinant plasmid is utilized as nucleic acid vaccine to perform intramuscular injection on the grass carps, and the nucleic acid vaccine enters the muscle cells and expresses VP35 protein of the grass carp reovirus in the muscle cells; thus, fish body immune cell proliferation is stimulated, antiviral related gene expression is up regulated, fish bodies are stimulated to generate antiviral antibodies, and capability of grass carps in resisting grass carp reovirus infection is effectively improved; furthermore, the grass carp reovirus S11 gene eukaryotic expression recombinant plasmid can be used for preventing a grass carp hemorragic disease caused by the grass carp reovirus in aquaculture.
Owner:HENAN NORMAL UNIV

Recombinant BHK cell line for stably expressing classical swine fever virus E0-E1-E2 protein, and applications of the same in preparation of vaccines and diagnosis reagents of classical swine fever

The present invention discloses a recombinant cell line for stably expressing classical swine fever virus E0-E1-E2 protein, and applications of the recombinant cell line in preparation of vaccines and diagnosis reagents of classical swine fever, wherein the recombinant cell line is BCSFV-E012, is preserved in the China General Microbiological Culture Collection Center, and has the preservation number of CGMCC No.7720. In addition, the present invention further discloses an establishment method for the cell line for stably expressing classical swine fever virus E0-E1-E2 protein, and a method for preparing a classical swine fever prevention vaccine composition by using the cell line. The present invention further discloses applications of the E0-E1-E2 protein stably expressed by the recombinant cell line in preparation of classical swine fever prevention vaccines and diagnosis reagents. The classical swine fever vaccine prepared by using the recombinant cell line has characteristics of high safety, good immunization effect, easy mass production, less being susceptible to exogenous virus pollution or influence of antibodies, and no classical swine fever virus non-structural protein antibody production so as to identify the vaccinated animal and the virus infected animal.
Owner:HARBIN WEIKE BIOTECH DEV +1

Pseudo-virus standard substance for nucleic acid diagnosis of novel coronavirus 2019-nCov and application thereof

PendingCN111378785AMeet the performance evaluation index requirementsOvercome the problem of CT value judgmentSsRNA viruses positive-senseMicrobiological testing/measurementGene synthesisBiological safety
The invention relates to a pseudo-virus standard substance for nucleic acid diagnosis of novel coronavirus 2019-nCov and application thereof. The pseudo-virus standard substance is prepared through the steps of gene synthesis, plasmid extraction, pseudo-virus packaging and the like. Wherein the synthesized gene comprises a novel coronavirus 2019-nCov gene RdRp, a Gene E and a Gene N; the 2019-nCovpseudo-virus standard substance is provided for the first time, is different from RNA synthesized in vitro and clinical positive living virus in the past, perfectly overcomes the defects of RNA and clinical positive living virus, is really suitable for performance evaluation of test kit, comprises limit of detection, specificity, repeatability and the like, and is further applied to clinical diagnosis of 2019-nCov; the pseudo-virus standard substance has the advantages of no pathogenicity, reproducibility, reliable quality control method and stable batch-to-batch, can be stably prepared and supplied for a long time, requires the biological safety level of P2 in a laboratory, and meets the safety requirements of many units.
Owner:仁宽(上海)生物科技有限公司

Method for constructing genetic engineering strains for producing (R)-acetoin and application of genetic engineering strains

The invention discloses a method for constructing genetic engineering strains for producing (R)-acetoin and application of the genetic engineering strains. The method includes optimizing codons of nucleotide sequences of alpha-acetolactate synthase genes, alpha-acetolactate decarboxylase genes and NADH (reduced form of nicotinamide adenine dinucleotide) oxidase genes and acquiring each gene cluster with three genes by the aid of artificial synthesis processes; inserting the gene clusters into expression vectors to obtain polycistron recombinant plasmids; introducing the polycistron recombinant plasmids into host bacteria E. coli and knocking out key genes of main byproduct synthesis paths to obtain the genetic engineering strains for producing the (R)-acetoin. The method and the application have the advantages that raw materials for the genetic engineering strains can come from wide sources and are low in cost, the strains are free of pathogenicity, oxidized form coenzymes NAD+ (nicotinamide adenine dinucleotide+) can be effectively regenerated, the strains are high in (R)-acetoin yield and production efficiency, the maximum yield can reach 72.1 g/L, and the optical purity can reach 99% at least; the (R)-acetoin is produced by the aid of non-grain cassava flour and inexpensive nitrogen sources which are used as fermentation raw materials, and accordingly the production cost can be reduced.
Owner:GUANGXI ACAD OF SCI

Method for preparing cyprini herpesvirus II antigen coated polyhedrosis based on baculovirus expression system

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.
Owner:苏州培恩特生物科技有限公司

Bacillus amyloliquefaciens and preparation method of exopolysaccharides thereof

The invention provides bacillus amyloliquefaciens EZ99. The bacillus amyloliquefaciens is gram positive bacteria, exists in plant rhizosphere soil, has no pathogenicity for people and plants, is collected from an orchard county, in the west of a Qilihe district in the Lanzhou city of the Gansu province, and has been identified, registered and preserved in the China common microorganism strain preservation center, and the preservation number is CGMCC No.13267. Component analysis is performed on bacterial colonies formed by the bacillus amyloliquefaciens on a PMA and MS culture medium, and it is found that the bacillus amyloliquefaciens has the characteristic of synthesizing and secreting exopolysaccharides. Optimal culture medium formulas and fermentation optimal conditions based on fermentation production of the exopolysaccharides of the strains are built, a fermentation solution is subjected to centrifuging, alcohol precipitation, dialysis and chromatographic separation, four purification polysaccharide components are obtained, and four-polysaccharide-component monosaccharide composition and monomer molecular weight are determined. The exopolysaccharides obtained through the strain and the method have the various applications of medicines, cosmetics, food additives, biological film raw materials and the like.
Owner:甘肃尚农生物科技有限公司
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