120 results about "Microorganism preservation" patented technology

The invention provides a method for permanently embedding an identity card number into a genome. The method comprises the following steps: permanently embedding an exogenous gene segment containing an identity card number into a genome by using CRISPR technology; and separating and reading specific DNA containing the identity card number. Thus, identity can always be identified via the permanently preserved DNA even an external packaging label is lost or wrong. The method is also applicable to preservation of microbes (including bacteria and viruses). For example, if a deactivated SARS virogene specimen is left, later generations can research on SARS viruses.

The present invention discloses a recombinant baculovirus strain rBac / PCV2Cap (microorganism preservation number: CGMCC NO.2083) efficiently expressing Porcine circovirus type 2 Cap protein and applications thereof. The recombinant baculovirus strain rBac / PCV2Cap constructed by the present invention can efficiently express recombinant PCV2-Cap protein in insect cells, and the expressed recombinant Cap protein, which has good immunocompetence and antigenicity, can serve as a subunit vaccine used to prevent the related plague caused by Porcine circovirus type 2 infection as well as a detecting and diagnostic antigen for Porcine circovirus type 2 serum antibody.

The invention provides a heat-resisting protective agent and an application thereof, and relates to the field of live microorganism deposit. The heat-resisting protective agent comprises the following substances: 40-80% of oligosaccharide, 3-12% of amino acid, 3-12% of gelatin, 3-12% of casein hydrolysate, 3-12% of polyvinyl pyrrolidone, 3-12% of glycerol, and 3-12% of poloxamer-68. The invention further provides the application of the heat-resisting protective agent in preparation of live vaccine freeze-dried powder. The heat-resisting protective agent is dissolved with a solvent; a protective agent solution is obtained; the protective agent solution is mixed with live virus liquid; a live vaccine is obtained; the live vaccine is freeze-dried; and then the live vaccine freeze-dried powder is obtained. The heat-resisting protective agent can improve heat resistance of the live vaccine, and the live vaccine freeze-dried powder prepared by the heat-resisting protective agent is stored for 2 months at 37 DEG C, and stored for 24 months at 15 DEG C.

The invention provides goose parvovirus (GPV) and applications thereof, and belongs to the technical field of biology. The invention provides a goose parvovirus GPV-YZ strain as well as applications thereof in preparing medicaments for preventing and treating the gosling plague disease. The microorganism preservation number is CGMCC NO.8160. The invention also provides a gosling plague disease viral vaccine as well as an anti-gosling plague yolk antibody. The goose parvovirus YZ strain provided by the invention has favorable immunogenicity for the present epidemic goose parvovirus, can resist toxicity attacking of the GPV-AV240 strain, the GPV-GD strain as well as the toxic strain of the goose parvovirus, and has the protection capability against the GPV-AV240 strain and the GPV-GD strain. After breeding geese are immunized by the gosling plague disease viral vaccine provided by the invention, young geese can be protected by maternal antibodies, and can be prevented from being infected by gosling plague viruses. The anti-gosling plague yolk antibody provided by the invention can prevent and treat gosling disease.

The invention provides a microbe-wilt bacterium preventing ANTI-8098A and the culture medium, culture method and biology application thereof. In particular a wilt bacterium preventing strain ANTI-8098A is provided, which is preserved in the Deutsche Sammlung von Mikroorganismen und Zellkulturen(DSMZ), with the preservation number being DSM ID01-100, the classification designation of the microbe being Bacillus cereus. The wilt bacterium preventing microbe collecting method, screen method, ferment production method and the culture medium thereof are provided, the supernatant containing the wilt bacterium preventing microbe or its ferment liquid, the biological pesticide and biological bactericide germicide containing its metabolic product are also provided. The prevention and cure effect of the wilt bacterium preventing microbe and the biological pesticide and biological bactericide germicide on the wilt bacteria of various kinds of plants in the field control experiment is over 80%, in addition, the quality and the output of the treated fruits are increased.

The invention discloses a transmissible gastroenteritis virus (H) attenuated vaccine strain and the application. The microorganism preservation number of the attenuated vaccine strain of the invention is CCTCC-V200609. The safety of attenuated strain TGEV of the invention is excellent, which has excellent immune protective rate. The attenuated strain of the invention can be applied in preparing diagnostic reagent for diagnosing transmissible gastroenteritis, and also can be applied in preparing single vaccine or mixed vaccine (active or inactivated vaccine) and the like.

The invention provides a fungus strain A. flavus G10 for degrading polyurethane plastic. The microorganism preservation number of the fungus strain A. flavus G10 is GDMCC 60537. The invention also provides a method for culturing the fungus strain A. flavus G10. The method includes the steps of isolating the fungus strain from the intestinal tract of a cricket; adopting PU as a unique carbon sourcefor culturing the fungus strain in a liquid culture medium to obtain a culture solution; diluting the culture solution and coating a solidified nutrient agar culture medium containing tetracycline antibiotics and a potato glucose agar culture medium with the culture solution to obtain cultured growing products. The cultured growing products are subcultured on fresh plates at 30 DEG C until a single fungus strain is obtained on each plate. The fungus strain A. flavus G10 can degrade the polyurethane plastic quickly.

The invention discloses a cell line ST / T7 capable of stably expressing T7RNA polymerase, the microorganism preservation number of which is: CGMCC No.24444. In the invention, a T7RNA polymerase gene is inserted into a eukaryotic expression vector pIRES2-EGFP to get the pIRES2-EGFP-T7 to transfect ST cells to obtain the cell line ST / T7 capable of stably expressing T7RNA polymerase. Through RT-PCR detection, expression plasmid containing a red fluorescent protein gene controlled by T7 promotors can be used to transfect the cell line so as to observe the expression of red fluorescent protein in a transcription product amplified form a ST / T7 cell to the T7RNA polymerase. The cell line ST / T7 can provide reverse T7RNA polymerase for use in reverse genetic manipulation of RNA-virus such as hog cholera virus, etc., and can be used as transcription and expression systems in vitro for research on genic structures and functions.

The present invention relates to microbe protecting technology, and is especially probiotics freeze drying process. The technological process includes inoculating yeast, faeces and lactobacillus in the amount of 8-10 % each, fermenting inside a tank at 37-39 deg.c temperature and 0.3-0.6 Kg / sq cm pressure, centrifuging the fermented product at 2000-2500 rpm to obtain bacterial mud, adding protecting agent to embed spawn, and freeze drying in a freeze drying machine to obtain freeze dried powder. The process is suitable for protection in short term and simple.

The invention belongs to the field of biological engineering and relates to a strain, in particular to a stenotrophomonas maltophilia and an application thereof. The stenotrophomonas maltophilia is preserved in the general microbiological center of Chinese microorganism preservation management committee on Mar. 20th, 2017 with the preservation number of CGMCC No. 13905. The stenotrophomonas maltophilia is used for degrading 2,7,11-westpac triene-4,6-diol and is applied in fermentation of a reconstituted-tobacco concentrate. During aging of tobacco leaves, the stenotrophomonas maltophilia can degrade 2,7,11-westpac triene-4,6-dialcohols into aromatic substances, and tobacco aroma is increased; in application of the reconstituted-tobacco concentrate, taste of reconstituted tobacco can be improved by improving aroma quality after conversion, increasing volume of aroma and reducing stimulation.

The invention discloses a porcine circovirus type 2 strain, an inactivated vaccine prepared from the porcine circovirus type 2 strain and application of the porcine circovirus type 2 strain, and belongs to the fields of isolation and application of the porcine circovirus type 2 strain. The invention firstly discloses a porcine circovirus type 2 Yh-1 strain, wherein the microorganism preservation number of the strain is CGMCC No. 10409. The invention also discloses a method for preparing a porcine circovirus preventing inactivated strain by virtue of the Yh-1 strain, wherein the method comprises the following steps: (1) amplifying the porcine circovirus type 2 Yh-1 strain and collecting a virus liquid; (2) adding an inactivating agent to inactivate the virus liquid and concentrating the virus liquid; (3) preparing an aqueous phase and an oil phase; and (4) mixing the aqueous phase and the oil phase, and emulsifying so as to obtain the Yh-1 strain. Results of animal experiments show that the inactivated vaccine prepared from the porcine circovirus type 2 Yh-1 disclosed by the invention is high in safety and is good in immunogenicity, and an immune protective rate reaches 100%; and the inactivated vaccine can offer complete protection over circovirus type 2 strong virulence attack and can be used for effectively preventing and controlling epidemic porcine circovirus.

The present invention discloses a hybridoma cell line and an anti-canine distemper virus N protein monoclonal antibody CDV-1N8 produced through the hybridoma cell line, and belongs to the field of CDV prevention and treatment. According to the present invention, the hybridoma cell line stably secreting the anti-CDV N protein monoclonal antibody is screened, wherein the microorganism preservation number is CGMCC No.8793, and experiment results prove that the monoclonal antibody CDV-1N8 secreted by the hybridoma cell line can specifically react with the CDV N protein, and does not react with the Vero cell protein; the monoclonal antibody can specifically recognize the CDV-N protein, and accurate positioning of the CDV-N protein B-cell epitope recognized by the monoclonal antibody is QITFLHSERS; and the monoclonal antibody CDV-1N8 and the CDV N protein virus-specific conservative B cell epitope polypeptide recognized by the monoclonal antibody can be made into the CDV infection diagnosis reagent so as to establish the foundation for establishment of the CDV serological differential diagnosis method.

The invention relates to a kit and method for extracting a microbial genome DNA by a magnetic bead method. The kit comprises a microbial preservation buffer solution, a bacteriolytic agent solution I, proteinase K, silica gel magnetic beads, isopropanol and an ethanol water solution, and further comprises a bacteriolytic agent II, a lysis adsorption solution and a reinforced scrubbing solution. The bacteriolytic agent II comprises MES, EDTA, Triton X-100, lysozyme and a lysozyme enhancer. The lysis adsorption solution comprises Tris-HCl, sodium chloride, EDTA, TrionX-100 and a nucleic acid separation and absorption promoter; and the reinforced scrubbing solution comprises Tris-HCl, lithium chloride and ethanol. By adopting the kit provided by the invention, cell walls of gram-positive bacteria and various fungi can be completely destroyed to form protoplasts, DNA of the protoplast cells can be rapidly released under the action of the lysis adsorption solution, the DNA is efficiently adsorbed on the silica gel magnetic beads, and after the DNA is washed by the scrubbing solution and eluted by an elution solution, microbial DNA with high concentration and purity can be obtained.

The invention relates to a recombination infectious haematopoietic necrosis virus (rIHNV HLJ-09) strain and a construction method and application thereof, and belongs to the field of biotechnologies. The microorganism preservation number of the rIHNV HLJ-09 strain is CGMCC No.8606, and the passage experiment of cells infected with recombinant virus shows that the virus can achieve passage stability. The rIHNV HLJ-09 is used in the intraperitoneal inoculation of juvenile rainbow trout, the result leads to the disease and death of the juvenile rainbow trout, and the lethallty rate can reach 90%, so that it shows that the virus has the similar pathogenic character as parental generation HLJ-09 strain virus. Furthermore, a rIHNV HLJ-09 reverse genetic system is used for replacing reporter gene green fluorescent protein (EGFP) encoding genes with NV ORF in the rIHNV HLJ-09, the virus is successfully rescued, and the rescued virus is named as rIHNV-EGFP. When a laser scanning confocal microscope is used for observing the cells infected with the rIHNV-EGFP virus, obvious green fluorescent can be observed, and it shows that the rIHNV HLJ-09 virus strain can be used for expressing foreign proteins.

The invention provides a compound biological preservative of food and a preparation method thereof, and relates to food preservation by microorganism. The biological preservative consists of nisin, wood vinegar and distilled water; and each 100 mL of the compound biological preservative of the food comprises 350 to 400 mg of the nisin, 15 to 25 mL of the wood vinegar, and the balance of the distilled water. The preparation method for the compound biological preservative comprises the following steps: mixing 350 to 400 mg of the nisin and 15 to 25 mL of the wood vinegar, and fixing the volume of the mixture to 100 mL by using the distilled water. The compound biological preservative of the food and the preparation method thereof comprehensively utilize the advantages and the characteristics of the nisin and the wood vinegar, and the nisin and the wood vinegar are compounded into the compound biological preservative of the food with oxidation resistance; and the compound biological preservative of the food has synergism, broad-spectrum preservation and the oxidation resistance; therefore, the compound biological preservative overcomes the defect that the prior food compound preservative is added with chemical agents and does not meet the requirement of the biological preservative basically, keeps the roast smoking flavor of the wood vinegar, and does not destroy the prior flavor of the food.

The invention discloses a preparation method of PCV2 (Porcine Circovirus2)-D. The microorganism preservation number of PCV2 (Porcine Circovirus2) is CGMCC (China General Microbiological Culture Collection Center) No.7245. According to the preparation method, the preference of genetic codon of the PCV2 is modified, and the preferred codon is modified into non-preference codon, so that the expressions of the components of the virus are reduced, further, the copy rate of the virus in host cells is lowered, the virus is weakened, and the PCV2-D is obtained by genetic modification. Furthermore, the proliferation speed of the PCV2-D is compared with that of wild type strains at a cellular level. The detection of PCR (Polymerase Chain Reaction) and IFA (Indirect Immunofluorescence Assay) prove that the PCV2-D can be infected and copied in cells, and has certain infection.

The invention discloses a bacillus subtilis zzl-002, preserved in the general microbiological center of Chinese microorganism preservation management committee (CGMCC for short, address: No.3, courtyard 1, Beichen road west, Chaoyang district of Beijing) in March 17, 2017, and the preservation number is CGMCC No.13758. The compound microorganism decayed bacterium is capable of shortening the fermentation time. The decayed organic fertilizer is capable of improving the fertility and reducing the plant diseases and insect pests.

The invention discloses an infectious laryngotracheitis recombinant virus strain for expressing a Newcastle disease virus F protein and establishment and the application of the strain and relates to the field of recombinant virus vaccines. Aiming at problems of recombinant virus establishment and the application of an exogenous immunogenic gene with a nonobligatory gene US9 locus of an ILTV (Infectious Laryngotracheitis Virus), the invention provides an infectious laryngotracheitis recombinant virus strain which is named as rILTV US9-NDV-F, the microorganism preservation number of the strain is CGMCC No.14738, and the virus strain is an infectious laryngotracheitis recombinant virus which is established by replacing a US9 gene of the infectious laryngotracheitis recombinant virus by the Newcastle disease virus F protein, establishing an acquirement deficiency US9 gene and by inserting an NDV-F (Newcastle Disease Virus-F Protein) expression box into a corresponding position of the gene,and is used for recombining the Newcastle disease virus F protein. The recombinant infectious laryngotracheitis recombinant virus strain disclosed by the invention is used for preparing vaccines forpreventing infectious laryngotracheitis and other chicken infectious diseases.

The application provides a strain of lactobacillus plantarum LP3 with a microorganism preservation number of CGMCC No. 14531. The bacterial strain can be used for fermented foods. The application alsoprovides a fructus lycii product prepared from a probiotics composition containing the bacterial strain, fructus lycii probiotics microecologics, a fructus lycii probiotics beverage and a preparationmethod thereof. The method is simple in operation, rich in source of raw materials, mild in preparation conditions, suitable for industrial production, the prepared fructus lycii product, the fructuslycii probiotics microecologics, the fructus lycii probiotics beverage make nutrients and probiotics in the fructus lycii compound and synergize, and preserve nutrients of the fructus lycii and probiotics; and in addition to original effects of the fructus lycii and probiotics, the characteristics of being easy to carry and eat, having a good taste and being easy to store are realized.

The invention provides a Bacillus megaterium Bacillus megaterium PP84. Deposition unit: China microorganism preservation management committee general microbiological center (CGMCC); address: Institute of Microbiology, No. 3, academy, No. 1, Beichen West Road, Chaoyang district, Beijing, China; deposition date: July 20, 2016; deposition number: CGMCC No. 12798. The Bacillus megaterium Bacillus megaterium PP84 has the advantages of having the capacity of using glyphosate as carbon and nitrogen sources; having the resistance capacity on heavy metals; being capable of decomposing and assimilating glyphosate, and adsorbing the heavy metals simultaneously; achieving the purpose of degrading the glyphosate in the environment.

The invention discloses bradyrhizobium japonicum vitrified cryopreservation liquid and a method for vitrified cryopreservation of bradyrhizobium japonicum and relates to microorganism vitrified cryopreservation liquid and a method for vitrified cryopreservation of microorganisms. The bradyrhizobium japonicum vitrified cryopreservation liquid contains ethanediol and is simple in compound composition. The bradyrhizobium japonicum vitrified cryopreservation liquid is composed of glycerinum, ethanediol, DMSO and distilled water. According to the method for vitrified cryopreservation of the bradyrhizobium japonicum, bradyrhizobium japonicum liquid is added into the bradyrhizobium japonicum vitrified cryopreservation liquid, and then the bradyrhizobium japonicum vitrified cryopreservation liquid is put into liquid nitrogen for fast vitrified cryopreservation. The bradyrhizobium japonicum vitrified cryopreservation liquid and the method for vitrified cryopreservation of the bradyrhizobium japonicum are used in the field of microorganism preservation.

The invention relates to a low-temperature microorganism preservation protectant, as well as a preparation method and use thereof. The low-temperature microorganism preservation protectant is prepared from the following components: non-reducing saccharide, glycerin, a nonionic surfactant, a potassium salt, glutamate and water. The invention also discloses a technical method for preserving bacillus calmette-guerin at low temperature for long. A technical support is provided for deep development of related researches on the bacillus calmette-guerin.

The invention discloses a porcine pseudorabies virus strain, an inactivated vaccine prepared from the porcine pseudorabies virus strain and an application of the porcine pseudorabies virus strain and belongs to the fields of separating and applying the porcine pseudorabies virus strain. The invention firstly discloses a porcine pseudorabies virus PRV-JL strain, and a microorganism preservation number of the porcine pseudorabies virus PRV-JL strain is CGMCC No.9903. The invention also discloses a method of applying the PRV-JL strain to preparing the porcine pseudorabies inactivated vaccine. The method comprises the following steps: (1) amplifying the porcine pseudorabies virus PRV-JL strain, and harvesting a virus liquid; (2) adding an inactivator to inactivate the virus liquid, and concentrating; (3) preparing a water phase and an oil phase; and (4) mixing the water phase and the oil phase, and emulsifying to obtain the porcine pseudorabies inactivated vaccine. Proved by animal experiment results that the inactivated vaccine prepared by the PRV-JL strain is high in safety, is good in immunogenicity, is high in immune protective rate, is capable of providing complete protection for both newly separated variant strain virulent strains and PRV MIN A strain virulent strains and is capable of effectively controlling current porcine pseudorabies.

The invention discloses a west nile virus NS1 protein monoclonal antibody, an identified B cell epitope thereof and application. A hybridoma cell strain which stably secretes anti-WNV (West Nile Virus)-NS1 protein monoclonal antibody is obtained by screening and has the microorganism preservation number of CGMCC No.4242. The monoclonal antibody WN-3D10 secreted by the hybridoma cell strain carries out specific reaction with WNV-NS1 protein but does not react with JEV (Japanese Encephalitis Virus)-NS1 protein. The monoclonal antibody and the WNV-NS1 protein virus specific B cell epitope identified by the monoclonal antibody can be used to be prepared into a reagent for identifying or diagnosing west nile virus and Japanese encephalitis virus, and lay a foundation for the serology differential diagnosis method for the WNV and Flavivirus virus.

The invention discloses an anti-glutamate dehydrogenase protein monoclonal antibody, a preparation method thereof and use thereof. In the invention, the antigenicity and hydrophobicity of the GDH gene of streptococci are analyzed, a pair of primers, after a signal peptide and a transmembrane domain are removed, is designed to clone the GDH gene, a GDH protein of WC-SS7 is successfully expressed by Escherichia coli, and a BALB / c mouse is immunized with the purified GDH recombinant protein to obtain a hybridoma cell line (with a microorganism preservation number: CGMCC No.3343) which can react with the GDH protein and stably secrete the antibody. The obtained GDH recombinant protein and the specific monoclonal can be used in researches on the structure and functions of Streptococcus suis, so a foundation is built for the method for serodiagnosis of clinic infection with Streptococcus suis. The antibody, the preparation method, and the use have great values in aspects such as further research on the function of the glutamate dehydrogenase in the pathogenic mechanism of Streptococcus suis and the establishment of a Streptococcus suis detection method.

The invention discloses a duck plague virus recombinant vaccine strain rDEVTK-EGFP for expressing enhanced green fluorescent protein (EGFP) genes and a constructing method and application of the duck plague virus recombinant vaccine strain rDEVTK-EGFP. The microorganism preservation number of the vaccine strain rDEVTK-EGFP is CGMCC No. 9456. According to the duck plague virus recombinant vaccine strain rDEVTK-EGFP, the constructing method and the application, a recombinant clone technology is adopted; a gene segment sCMV-EGFP containing enhanced green fluorescent proteins (EGFP) and sCMV promoter sequences is inserted into a duck plague virus TK gene region; recombinant EGFP duck plague viruses with an sCMV-EGFP expression cassette inserted into the corresponding position of a TK gene is constructed; the EGFP genes are stably expressed by the recombinant viruses. The invention further relates to a method for constructing the recombinant duck plague virus vaccine strain for stably expressing other poultry pathogeny exogenous genes and the application of the recombinant duck plague virus vaccine strain to preparation of vaccines for preventing duck plagues and other poultry infectious diseases.

The invention discloses a preparation method of a resin adsorbent based on composite kieselguhr. The kieselguhr is adopted as a first carrier, a resin is adopted as a second carrier, and a microorganism strain is fixed. The resin is prepared through the steps of adding 5 to 20 parts by weight of kieselguhr immobilized microorganism, 0.1 to 0.5 part by weight of a coupling agent, 1 to 3 parts by weight of butyl hydroxy anisd and 1 to 5 parts by weight of tetrahydrocannabinol into an acrylate copolymer emulsion, uniformly stirring at the normal temperature at low speed, and vacuum drying to obtain the resin adsorbent based on the composite kieselguhr. The adsorbent prepared through the method has the advantages of fast oil absorption speed, high oil absorption capacity, long microorganism preservation time, long service life and the like.

The invention provides a manufacturing method of a loess bed formed by a loess block and the loess bed manufactured with the method. According to the manufacturing method of the loess bed formed by the loess block, the loess block used for manufacturing the loess bed is dried in a natural state instead of in a high-temperature state, and then various useful components and microorganisms in loess are perfectly preserved, the loess is converted into powder and then is stirred and is dried in a natural mode in a forming framework, and the surface of the loess block is enabled to be smooth, and the loess powder is prevented from flowing.

The invention relates to cultivation of microorganisms, and particularly relates to photosynthetic bacteria SC01 as well as a cultivation method and an application thereof. The photosynthetic bacteria are Rhodobacter sphaeroides, and preserved in China Microbiological Collection Center on September 25, 2013, with the preservation number of CGMCC No.8248. The photosynthetic bacteria SC01 are used in a cultivation process of breeding organisms as an additive so as to increase the food intake of breeding organisms, and reduce the residual feed rate. The bacterial strain provided by the invention has the characteristics that a screening method is easy and feasible, the production, storage and transportation are convenient, the using effect is obvious and the like, and has good application prospect.