37results about How to "Good passage stability" patented technology

The invention provides a Vero cell line capable of stably expressing protein P54 of an African swine fever virus. The collection number of the cell line is CGMCC No.14316. The cell line not only can identify a polyclonal antibody of African swine fever, but also can generate specific reaction with positive serum of an African swine fever-infected pig. The monoclonal Vero cell line of an African swine fever P4 antigen can be used for stably expressing the protein P54 of the African swine fever virus, and can be used as a cell model or a virus infection model; the cell transmission stability ishigh, the growing speed is high, and culture conditions are simple; a large number of cell lines can be provided; the cell line has a good application prospect for researches on detection and pathogenesis mechanisms of the African swine fever.

The invention discloses a three-dimensional simulation culture system for stem cells, and belongs to the technical field of biology. The invention solves the problems of complex operation, low efficiency, small yield and poor quality in an existing in-vitro culture of stem cells. The three-dimensional simulation culture system for the stem cells comprises a stem cell nest, a heart region first peristaltic pump, a lung region, a kidney region, a spleen region, a stomach region, a liver region, a first fluid pressure gauge, a first pressure regulating valve, a liquid level meter, a three-way valve, a first control valve, a temperature regulator, a thermometer, an oxygen / carbon dioxide measuring instrument, a PH measuring instrument, a nutritional ingredient measuring instrument, a second control valve, a second peristaltic pump, a third peristaltic pump, a urea / uric acid / inorganic salt measuring instrument, a fourth peristaltic pump, a third control valve, a second fluid pressure gauge,a second pressure regulating valve, a fourth control valve, a fifth control valve, a fifth peristaltic pump, a sixth control valve, a gas permeable membrane, a dialysis membrane, a central control platform, a cloud data center and an electromagnetic field. According to the three-dimensional simulation culture system for the stem cells, the stem cells with higher quality and quantity can be cultured, the stability is good, and the treatment effect of the stem cells is improved.

The invention discloses a caproic acid fermented strain and a caproic acid extracting method. The caproic acid fermented strain is js-22-03, js-22-07 and js-22-12. Bred bacterial strains are good in passage stability, and the yield of the bred bacterial strains is increased to 1.90 g / 100 mL from 1.20 g / 100 mL compared with an original bacterial strain. The high-yield bacterial strains js-22-03, js-22-07 and js-22-12 are obtained through culture and screening, and the advantages of being short in fermentation time, stable in genetic property, high in yield and the like are achieved. The fermentation time is shortened to be within 10 days, and the caproic acid yield is increased to 1.90 g / 100 mL from original 1.20 g / 100 mL.

The invention discloses screening and application of a bacillus strain for producing cellulase, and belongs to the technical field of bioengineering. According to the invention, a strain of bacillus subtilis is screened from animal waste, the activity of cellulase produced by fermentation of the bacillus subtilis on a shake flask level reaches 118.67 U / mL, and the bacillus subtilis has good passage stability. The method for producing the cellulase by using the bacillus subtilis is simple and feasible in process operation, low in culture medium cost and suitable for industrial production.

The invention discloses streptomyces sp. for producing epsilon-polylysine and an application of the streptomyces sp., and belongs to the field of biotechnology. A streptomyces albulus (streptomyces albulus) M-Z18 bacterial strain is preserved in the China General Microbiological Culture Collection Center, and the preservation number is CGMCC NO: M 2019589. The streptomyces albulus M-Z18 bacterialstrain can quickly grow on a culture medium using succinic acid as a carbon source, and has the capacity for high efficiency synthesis of epsilon-polylysine having sulfaguanidine resistance. The bacterial strain is subjected to fed-batch fermentation under a pH impact strategy, the accumulation concentration of the epsilon-polylysine can achieve 54.7g / L, the production efficiency can achieve 6.83g / L / d, and the streptomyces sp. is an epsilon-polylysine industrial production bacterial strain having great potential.

The invention discloses streptomyces albulus (GS-114) and a method of the same to prepare epsilon-polylysine, and belongs to the field of microbial fermentation engineering. The provided streptomycesalbulus (GS-114) has resistance to streptomycin, gentamicin and rifamycin with 3 [mu]g / mL, 1 [mu]g / mL and 0.1 [mu]g / mL or higher concentrations, and can efficiently synthesize epsilon-PL in quantity,the yield with 192 h of fermentation can reach 56.3 g / L, and production efficiency can reach up to 7.03 g / L / d. The streptomyces albulus (GS-114) also has good passage stability and has strong potential for the industrial production of the epsilon-PL.

The invention provides a BHK-21 cell line for stably expressing schmallenberg virus (SBV) nucleocapsid protein. The preservation number of the BHK-21cell line is CGMCC NO. 7616, and the cell line is named BHK-21-pLV-EGFP-SBV-N. The cellular immunity fluorescence experiment result shows that the cell line can not only have specific reaction with a monoclonal antibody 2C8 of SBV, but also can recognize positive serum of SBV infected cattle. The BHK-21 cell line for stably expressing the SBV N protein can be used for detecting schmallenberg disease and has good application prospect.

The invention discloses a recombinant plasmid for genetic transformation of pleurotus eryngii and application thereof and provides a DNA (Deoxyribonucleic Acid) segment and the recombinant plasmid containing the DNA segment, wherein the DNA segment sequentially comprises a promotor shown as a sequence 1 and a terminator shown as a sequence 2 from upstream to downstream. According to a method for genetic transformation of the pleurotus eryngii, which is established by the invention, the foundation for theoretical basis and technical support for fundamental research and molecular breeding of the growth and development mechanism of the pleurotus eryngii is laid, the storage for developing a novel bioreactor is provided and wide application field for developing and utilizing pleurotus eryngiiresources and bioactive substances thereof is opened up.

The invention provides an L-homoserine production bacterial strain. The L-homoserine production bacterial strain is characterized by containing a single and stable chromosome and not containing DNA carrier plasmids or other free DNA carriers, one or more genes related to L-threonine synthesis on the chromosome DNA of the bacterial strain are subjected to knockout or weakening, and / or the genes aresubjected to mutation for enhancement, and / or one or more genes related to the L-homoserine metabolic pathway are integrated and copied on the chromosome DNA of the bacterial strain.

The invention provides lactobacillus paracasei SLP16 for producing polysaccharides, application of the lactobacillus paracasei SLP16 and a feed additive prepared from the lactobacillus paracasei SLP16, and relates to the technical field of feed additives. The lactobacillus paracasei SLP16 disclosed by the invention is Lactobacillus paracasei SLP16; the Latin name is Lactobacillus paracasei, the name of the preservation unit is Guangdong Microbial Culture Collection Center; the address is Floor 5, Building 59, No. 100 courtyard, Xianlie Middle Road, Guangzhou city; the preservation date is July 5, 2021; and the preservation number is GDMCCNo: 61769. The SLP16 disclosed by the invention is still stable after passage, can inhibit escherichia coli and staphylococcus aureus, has good cholate-resistant and acid-resistant effects, has cohesiveness, hydrophobicity and oxidation resistance, and also can generate exopolysaccharides.

The invention relates to 'a subcloned cell strain TF-1-A2, a preparation method thereof and application' and belongs to the technical field of biology. The preparation method of the stable subcloned cell strain TF-1-A2 includes the steps: domesticating GM-CSF (granulocyte-macrophage colony-stimulating factor) dependent TF-1 cells into NGF (nerve growth factor) dependent strains; cloning the induced NGF dependent TF-1 cells by the aid of methylcellulose semisolid media; picking monoclonal cells from the semisolid media and enlarging and cultivating the monoclonal cells; and preparing a dose-effect curve of each monoclonal cell strain to the NGF and performing screening to obtain the subcloned cell strain TF-1-A2. The subcloned cell strain TF-1-A2 obtained by the preparation method is highly sensitive to the NGF and has fine reactivity and continuous inheritance stability. A TF-1 cell growth method established by the aid of the subcloned cell strain TF-1-A2 is used for externally and quantitatively measuring biological activity of the nerve growth factor, and is high in signal-to-noise ratio, fine in repeatability and superior to an existing method.

The invention provides a method for rapid preparation of epidemic and infectious bronchitis vaccine. The method takes infectious clone of an infectious bronchitis virus H120 vaccine strain as a skeleton carrier, then replaces an antigen gene in the skeleton carrier with a targeted antigen gene of an infectious bronchitis epidemic virus strain, so as to obtain the method of recombining bronchitis virus, and the targeted antigen gene is prepared from S gene fragments of infectious bronchitis epidemic strains with different serotypes / genotypes in an gomphosis mode; replacement can also be the simultaneous replacement of the targeted antigen gene and a N gene, and a signal peptide region of an original S1 gene in the skeleton carrier needs to be preserved during the replacement. The method forthe rapid preparation of the epidemic and infectious bronchitis vaccine has the beneficial effects that an operation method is simple and easy to conduct, therepeatability is high, the generation stability is good, the frequent variation of IBV epidemic can be quickly and efficiently tackled, and the method provides a new idea for the construction of a carrier vaccine.

The invention discloses an aspergillus niger strain and application thereof in ochratoxin A degradation, and belongs to the field of gene engineering. The invention provides the Aspergillus niger FS-UV-21 strain which is preserved in the China General Microbiological Culture Collection Center on November 9, 2020, and the preservation number of the Aspergillus niger FS-UV-21 strain is CGMCC No. 20751. The aspergillus niger FS-UV-21 has a good degradation effect on ochratoxin A, the degradation rate reaches 74.49%, and the degradation rate is improved by 18.23% compared with that of an original strain; the time for achieving the optimal degradation effect is advanced and is shortened by 18 hours; aspergillus niger belongs to food-grade microorganisms, has relatively high food safety, and can realize efficient biological degradation of fungaltoxin.

The invention relates to a strain of streptomyces albidus AAE89, which is named as Streptomyces albidus AAE89 in Latin. The strain is preserved in China General Microbiological Culture Collection Center on October 12, 2020, the preservation address is No.3, Yard 1, Beichen West Road, Chaoyang District, Beijing, and the preservation number is CGMCC 20880; the streptomyces albidosus AAE89 provided by the invention has stronger acid resistance, and compared with an original strain, the streptomyces albidosus AAE89 has the advantages that the dry weight of a thallus is increased by 46.92% after the streptomyces albidosus AAE89 is cultured for 48 hours under the condition that the initial pH is 4.0, and the survival rate of the thallus is increased by 1.81 times after the thallus is stressed for 48 hours under the condition that the pH is 3.0.

The invention discloses a high-yielding penicillin strain, a screening method of the strain, and application of the strain in penicillin production, and belongs to the technical field of antibiotics production and application. The strain is characterized in that penicillin fermenting unit is increased by 13.2%, so that the yield of penicillin is effectively increased, and the production cost is decreased. The screening method comprises the following steps: (A) preparing a protoplast; (B) performing high-energy electronic mutagenesis; (C) rationally screening; and (D) screening with a shake bottle. The penicillin strain screening method is high in purposiveness, and high in mutation probability.

The invention discloses a strain for highly yielding enramycin, simultaneously provides a screening method of the strain, and provides application of the strain in enramycin production. The preserving number of the strain DCS067 for highly yielding enramycin is CGMCCONO: 6220, the preserving unit is China General Microbiological Culture Collection Center (CGMCC), and the preserving date is June 14, 2012. The DCS067 strain enables production rate of enramycin to be improved by 47.80%.

The invention discloses a strain capable of highly producing vitamin K2 (MK-7), a screening method of the strain and a method for producing the vitamin K2 (MK-7) by using the strain, and belongs to the field of microbial fermentation. The strain is preserved in China Center for Type Culture Collection on August 23, 2021, and the preservation number is CCTCC M20211066. Mutagenesis is performed by using nitrosoguanidine, and the strain is obtained through shake flask screening, and has the characteristics of high shake flask titer, good passage stability and the like. When the strain is used for fermentation, a fermentation unit can reach 240 mg / L or above in cooperation with an optimal fermentation culture medium and a control process, the fermentation unit is improved by 243% compared with that of an original strain, the yield of the vitamin K2 (MK-7) can be effectively increased, the cost of raw materials, water, electricity, steam and the like for fermentation is obviously reduced, pollution emission of the unit yield is reduced, and the strain has high economic benefits.

The invention discloses a strain of streptomyces producing ε-polylysine and its application, belonging to the technical field of bioengineering. The Streptomyces albulus strain (Streptomyces albulus) M-Z18 of the present invention has been deposited in the China General Microorganism Collection and Management Center on July 29, 2019, and the preservation number is CGMCC NO: M 2019589. The Streptomyces albicans M-Z18 strain of the present invention can grow rapidly on a medium with succinic acid as a carbon source, and has the ability to efficiently synthesize ε-polylysine with sulfaguanidine resistance. The strain of the present invention carries out fed-batch fermentation under the pH shock strategy, the cumulative concentration of ε-polylysine can reach 54.7g / L, and the production efficiency can reach 6.83g / L / d, which is a strain of ε-polylysine with great potential Polylysine industrial production strain.

The invention relates to an RAW264.7 single-cell stable cell line transfected with red fluorescent protein and a screening method thereof. The screening method includes: (1) adding pDsRED2-C1 plasmidand a transfection reagent into RAW264.7 cells for transfection; (2) applying screening pressure to the cells obtained in step (1); and (3) according to a fluorescence observation result, selecting the positively transfected cells obtained in step (2) for digestion, resuspending, single-cell dilution plate paving, and conducting single-cell stable cell line screening. The method for screening RAW264.7 single-cell stable cell line transfected with red fluorescent protein can acquire a RAW264.7 single-cell cell line with good transfection effect, and the cell line has good passage stability.

The invention provides a streptomyces fungicidious mutant strain DC461, wherein the collection number is CGMCC (China General Microbiological Culture Collection Center) No. 5811, the collection unit is China Committee for Culture Collection of Microorganisms, and the collection date is February 27, 2012. The invention further provides a preparation method of the strain and application of the strain in terramycin production. The mutant strain DC461 provided by the invention is bearable to a high-concentration defoamer and can be used for increasing the production level of terramycin.

The invention discloses a cell line for detecting the activity of human interferon-kappa and a construction and detection method thereof. The cell line is obtained by transfecting eukaryotic cells with ISRE (interferon-sensitive response element) luciferase reporter genes and stably screening monoclonal cell lines. The cell line constructed by the invention is easy to culture and amplify and good in passage stability, and has the advantages of high sensitivity, short detection time, completion within 4-6 hours, and capability of greatly improving the activity release efficiency of the human interferon-kappa in the production process.

The invention provides a production strain of L-homoserine, which is characterized in that: it has a single stable chromosome, does not contain plasmid form or other free DNA carriers, and one or more of its chromosomal DNA is related to the synthesis of L-threonine The genes of the patients are knocked out or weakened, and / or mutated to enhance, and / or one or more genes related to the L-homoserine metabolic pathway are integrated on their chromosomal DNA.

The invention discloses a recombinant plasmid for genetic transformation of pleurotus eryngii and application thereof and provides a DNA (Deoxyribonucleic Acid) segment and the recombinant plasmid containing the DNA segment, wherein the DNA segment sequentially comprises a promotor shown as a sequence 1 and a terminator shown as a sequence 2 from upstream to downstream. According to a method for genetic transformation of the pleurotus eryngii, which is established by the invention, the foundation for theoretical basis and technical support for fundamental research and molecular breeding of the growth and development mechanism of the pleurotus eryngii is laid, the storage for developing a novel bioreactor is provided and wide application field for developing and utilizing pleurotus eryngiiresources and bioactive substances thereof is opened up.

The invention discloses a method for simultaneously preparing two kinds of monoclonal antibodies. In the method, the conjugate of 33F type pneumonia polysaccharide and hepatitis B surface protein is used as an antigen to immunize mice, and mouse splenocytes with higher antibody levels after immunization are selected. Fusion with SP2 / 0 myeloma cells, and then specific screening by 33F pneumonia polysaccharide, hepatitis B surface protein, and 33F pneumonia polysaccharide and hepatitis B surface protein conjugates as antigens to obtain hybridoma cells, and at the same time prepare a specific recognition of 33F pneumonia Monoclonal antibodies to polysaccharide and hepatitis B surface protein. The method of the present invention to simultaneously prepare monoclonal cell line cells and specific monoclonal antibodies for polysaccharides and proteins after immunization with pneumonia conjugates using hepatitis B surface protein as a carrier saves workload and improves screening efficiency. At the same time, the present invention The two monoclonal antibodies provided have the advantages of high specificity, high sensitivity and good passage stability.

The invention provides a Vero cell line capable of stably expressing protein P54 of an African swine fever virus. The collection number of the cell line is CGMCC No.14316. The cell line not only can identify a polyclonal antibody of African swine fever, but also can generate specific reaction with positive serum of an African swine fever-infected pig. The monoclonal Vero cell line of an African swine fever P4 antigen can be used for stably expressing the protein P54 of the African swine fever virus, and can be used as a cell model or a virus infection model; the cell transmission stability ishigh, the growing speed is high, and culture conditions are simple; a large number of cell lines can be provided; the cell line has a good application prospect for researches on detection and pathogenesis mechanisms of the African swine fever.

The invention discloses bacillus with relatively high tolerance to methanol and belongs to the technical fields of microbial culture, molecular biology and application. The strain disclosed by the invention has been preserved in the China Center for Type Culture Collection on April 6, 2021, and the preservation number is CCTCC NO: M 2021326. The strain can tolerate 20% or even higher methanol, has good passage stability, still keeps methanol tolerance after 50 generations of passage, can be applied to construction of methylotrophication strains and synthesis of high-added-value products by using methanol as a substrate, and has wide industrial application prospects.

The present invention provides a BHK-21 cell line stably expressing Schmallenberg virus (SBV) nucleocapsid protein, its preservation number is CGMCC No.7616, named BHK-21-pLV-EGFP-SBV-N . The results of cell immunofluorescence test showed that the cell line not only reacted specifically with SBV monoclonal antibody 2C8, but also recognized the positive serum of SBV-infected cattle. The BHK‑21 cell line stably expressing SBV N protein can be used for the detection of Schmallenberg disease and has a good application prospect.

The present invention provides a kind of penicillin V potassium high-yielding bacterium and its application in fermentative production of penicillin V potassium, the penicillin V potassium high-yielding bacterium is Penicillium chrysogenum ( Penicillium Chrysogenum ) B181126 strain, the preservation date is October 14, 2019, the preservation number is CCTCC NO: M2019823, and the preservation unit is the China Center for Type Culture Collection (CCTCC). When Penicillium chrysogenum B181126 is used in production, with the optimal fermentation process, its fermentation unit can reach more than 80,000 U / mL, which is 36.4% higher than the starting strain, and the fermentation unit of penicillin V potassium is greatly increased. At the same time, 4 ‑ Hydroxypenicillin V potassium impurity content is less than 1.2%.