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43results about How to "Avoid Biosafety Hazards" patented technology

Clostridium perfringens Beta toxin recombination subunit vaccine and production method thereof

ActiveCN109078178AEfficient expressionEfficient soluble expressionAntibacterial agentsBacterial antigen ingredientsClostridium perfringens beta toxinVaccine Production
The invention relates to a clostridium perfringens Beta toxin recombination subunit vaccine and a production method thereof. The prepared clostridium perfringens Beta toxin recombination subunit vaccine is produced by using a recombination clostridium perfringens Beta toxin protein which is processed by codon optimization and contains 4 amino acid mutations, namely integrity and spatial conformation of a natural toxin protein are reserved in the greatest degree, so immunogenicity of the natural toxin protein is kept, and a biological potential safety hazard caused by the single amino acid mutation is avoided. The vaccine further has the advantages of simple preparation process, low immunizing dose, good vaccine efficacy and the like. Compared with a current commercial clostridium perfringens natural toxin inactivated vaccine in China, a bio-safety risk in a vaccine production process is greatly reduced. The vaccine is a perfect candidate vaccine for updating a current C-type clostridium perfringens toxin vaccine in China. In addition, while a mixed vaccine is prepared by the vaccine with other antigens, the mixed vaccine can be prepared without increasing a using dose of the mixedvaccine.
Owner:CHINA INST OF VETERINARY DRUG CONTROL

Chicken interferon alpha biological activity detection method

The invention discloses a chicken interferon alpha biological activity detection method, and applications thereof. The chicken interferon alpha biological activity detection method comprises followingsteps: PCR amplification is adopted to obtain Mxp gene segments of chicken Mx protein; pCMV of pEGFP-N1 vector plasmid is removed; the Mxp gene segments of chicken Mx protein obtained through PCR amplification are subjected to construction of pEGFP-N1-Mxp plasmid through replacing of pCMV of orginal pEGFP-N1 vector plasmid with T4DNA ligase; the pEGFP-N1-Mxp plasmid is adopted for cell transfection, and a cell strain capable of realizing stable transfection is obtained through screening using neomycin; the screened cell strain capable of realizing stable transfection is subjected to cloning culture, chicken interferon alpha is added for co-incubation with the cell strain which is capable of realizing stable transfection and is treated throug cloning culture, so that Mx gene promoter activity is activated to promote intracellular EGFP expression, the intensity of fluorescence emitted by cells after excitation light source irradiation is positively related to chicken interferon alpha biological activity, so that quantitative evaluation of chicken interferon alpha biological activity is realized.
Owner:ANHUI JIUCHUAN BIOTECH

Method for producing beta-mannase by adopting lactic acid bacteria cultured by taking konjaku flour as carbon source

The invention provides a method for producing beta-mannase by adopting lactic acid bacteria cultured by taking konjaku flour as the carbon source, relating to a method for producing beta-mannase by adopting lactic acid bacteria, for solving the problems that during the producing of mannase by adopting fungi in the prior art, the fermentation period is long, the bacteria for producing mannase has the potential biological safety hazards, and the activity of the lactic acid bacteria in producing the mannase is low. The method comprises the following steps: 1, inoculating lactobacillus plantarum to an MRS solid medium panel, and carrying out culturing; 2, picking single colonies, and carrying out culturing in MRS liquid medium; 3, carrying out centrifugalization, abandoning the supernatant, collecting thalli, and adjusting the cell concentration, thus obtaining the seed solution; 4, inoculating the seed solution to a fermentation medium, thus obtaining fermentation liquor; and 5, centrifugalizing the fermentation liquor, abandoning the thalli, and taking the supernatant, thus obtaining the mannase crude enzyme. The crude enzyme produced by adopting the method provided by the invention is high in enzyme activity, low in production cost, and short in production period, and the produced bacterial strain has no biological safety hazards. The method is applied to the field of microbial fermentation enzyme production.
Owner:蔡河齐

Detection method for biological activity of porcine interferon alpha through luciferase reporter gene method

The invention discloses a detection method for the biological activity of a porcine interferon alpha through a luciferase reporter gene method and application of the detection method. The method comprises the following steps: obtaining a gene segment of pMx1 of porcine Mx1 protein by adopting PCR (Polymerase Chain Reaction) amplification; inserting the gene segment of the pMx1 into a 5' end of a pGL3-basic vector luc gene; then carrying out the PCR amplification to obtain a pMx1-luc fused gene segment; replacing gene segments of pCMV (Porcine Cytomegalovirus) and EGFP (Enhanced Green Fluorescent Protein) in a pEGFP-N1 vector by utilizing the pMx1-luc fused gene segment; constructing a pMx1-luc plasmid and transfecting a cell; selecting a stably-transfected cell line; carrying out clonal culture on the screened stably-transfected cell line; preparing a standard curve by taking a porcine interferon alpha standard product which is subjected to gradient dilution; adding a porcine interferon alpha sample to be detected into cells subjected to the clonal culture for co-incubating; then detecting the fluorescence intensity and combining the standard curve to evaluate the valence of the porcine interferon alpha sample to be detected.
Owner:ANHUI JIUCHUAN BIOTECH

Nontoxic clostridium perfringens and clostridium septicum fusion protein vaccine and production method thereof

The invention relates to a nontoxic clostridium perfringens and clostridium septicum fusion protein vaccine and a production method thereof. The fusion protein vaccine prepared by the method adopts aclostridium perfringens epsilon toxin which is subjected to codon optimization and contains three amino acid mutations, and spoilage which contains four amino acid mutations and eleven amino acid deletions, namely not only kept the integrity and spatial conformation of the natural toxin are to the maximum extent, but also the biological safety hazard caused by single amino acid mutation is avoided. The vaccine also has the advantages of simple preparation process, excellent vaccine efficacy and the like, greatly reduces the biological safety risk in the vaccine production process. Compared with the current commercialized sheep fast-epidemic and sheep enterotoxemia inactivated vaccines in China, the vaccine is an ideal candidate vaccine for upgrading existing clostridium perfringens and clostridium septicum vaccines in China; in addition, when combined vaccines are prepared by the vaccine together with other antigens, the combined vaccine can be prepared without increasing the use doseof the combined vaccine.
Owner:CHINA INST OF VETERINARY DRUG CONTROL

Centrifuge tube with double-layer structure

The invention relates to a centrifuge tube with a double-layer structure. The centrifuge tube comprises an inner pipe and an outer pipe, the inner pipe is provided with a first internal cavity; the outer pipe is provided with a second inner cavity; the outer pipe is sleeved outside the inner pipe; an annular sealing ring is arranged between the outer pipe and the inner pipe, the annular sealing ring is fixed to the outer pipe and used for achieving sliding sealing, the inner pipe is provided with a communicating hole used for communicating the first inner cavity with the second inner cavity, the outer pipe is provided with a sealing piece used for sealing the communicating hole, and the outer pipe is used for moving from a first position to a second position relative to the inner pipe; theinvention provides the centrifugal tube with the double-layer structure, the supernatant can be conveniently and safely separated without pollution after the centrifugation is completed, and the supernatant can be effectively prevented from being dumped to disturb the sample gathered on the lower layer so as to pour out the precious and rare cell components along with the supernatant, and more importantly, the separated supernatant can be effectively stored and isolated so as to effectively prevent the biological pollution and effectively eliminate the biological safety hidden danger.
Owner:SICHUAN CANCER HOSPITAL

Method for detecting drug resistance of antituberculosis drugs based on overlap-extension PCR

The invention relates to a method for detecting the drug resistance of antituberculosis drugs based on an overlap-extension PCR. By adopting an overlap-extension PCR method, four drug-resistance related segments of rpoB, embB and katG genes and an inhA promoter are connected and amplified into a fusion segment with the length of about 700 bp in a single tube, drug-resistance mutation information of the four segments is obtained just through a one-time sequencing reaction, and accordingly the condition of drug resistance of some kind of mycobacterium tuberculosis to three kinds of first-line antituberculosis drugs is known. If the method is applied and popularized clinically, the detection cost is greatly reduced, the economic burden of a patient is reduced, and the good social benefit is achieved. In short, by means of the method that the single-tube PCR amplification rpoB-embB-katG-inhA fusion segment is used for sequencing, so that the drug resistance condition of three kinds of first-line antituberculosis drugs is detected, the economic burden is reduced for detection of clinical drug resistance tuberculosis, the detection accuracy is improved, and the method is easy and convenient to implement, accurate and low in cost.
Owner:重庆市公共卫生医疗救治中心

Detection method for biological activity of sheep interferon tau through luciferase reporter gene method

The invention discloses a detection method for the biological activity of a sheep interferon tau through a luciferase reporter gene method and application of the detection method. The method comprisesthe following steps: obtaining a gene segment of pMx1 of sheep Mx1 protein by adopting PCR (Polymerase Chain Reaction) amplification; inserting the gene segment of the pMx1 into a 5' end of a pGL3-basic vector luc gene; then carrying out the PCR amplification to obtain a pMx1-luc fused gene segment; replacing gene segments of pCMV (Porcine Cytomegalovirus) and EGFP (Enhanced Green Fluorescent Protein) in a pEGFP-N1 vector by utilizing the pMx1-luc fused gene segment; constructing a pMx1-luc plasmid and transfecting a cell; selecting a stably-transfected cell line; carrying out clonal cultureon the screened stably-transfected cell line; preparing a standard curve by taking a sheep interferon tau standard product which is subjected to gradient dilution; adding a sheep interferon tau sampleto be detected into cells subjected to the clonal culture for co-incubating; then detecting the fluorescence intensity and combining the standard curve to evaluate the valence of the sheep interferontau sample to be detected.
Owner:WUHU YINGTE FEIER BIOLOGICAL PROD IND RES INST CO LTD

Immunosuppressive myocarditis mouse model and construction method and application thereof

The invention provides an immunosuppressive myocarditis mouse model and a construction method and application thereof. The method comprises the following steps that a male BALB/c mouse of 4-6 weeks old is selected as an experimental mouse for constructing the myocarditis mouse model, and observation and feeding are conducted for three days; after feeding is conducted for three days, a cyclosporine A solution with the preset concentration is injected in an intraperitoneal injection mode, the injection amount each time is 40-50mg/kg of the mouse weight, injection is conducted once every other day, and injection is conducted three times in total; and after the last injection is conducted for one day, the myocarditis mouse model is obtained. According to the immunosuppressive myocarditis mouse model and the construction method and application thereof, cyclosporine A serves as an immunosuppressor, myocarditis is successfully induced according to a specific administration method, myocardial damage is caused, the myocarditis mouse model is constructed, and death does not exist; compared with a mouse model constructed through virus infection and LPS induced inflammation in the prior art, the immunosuppressive myocarditis mouse model has the advantages of being safe and effective, and the tolerance of the experimental mouse is better.
Owner:HUNAN UNIV OF CHINESE MEDICINE
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