61results about How to "Good genetic stability" patented technology

The invention relates to the technical field of gene engineering, and discloses a KASP marker related to thousand seed weight of wheat and application of the KASP marker. The SNP locus is located at the 33397938bp position of the wheat 1A chromosome, and the polymorphism is C / T. Through phenotypic difference and exon capture analysis, the SNP site closely related to the thousand seed weight is determined, and a specific KASP primer combination is designed, so that rapid and accurate identification of the wheat thousand seed weight is realized. The application of the technology has significant significance in accelerating high-yield breeding of wheat. According to the method, the selection efficiency is improved, the breeding period is shortened, and wheat varieties with ideal thousand seed weight can be quickly identified and cultivated. By using the KASP primer combination, an effective tool is provided for molecular marker-assisted selection of wheat yield-related traits, and the overall yield and quality of crops can be improved.

The invention relates to the technical production field of metabolic engineering and genetic engineering, and particularly provides a uracil production strain as well as a construction method and application thereof. According to the strain, a uridine kinase gene udk is knocked out on an E.coli W3110 genome, and meanwhile, a T7RNA polymerase gene is integrated; the uridine monophosphate kinase gene pyrH is weakened; the method comprises the following steps: overexpressing a carbamyl phosphate synthetase gene carAB, a ribose phosphate pyrophosphate kinase gene prsA, a pyrimidine-5 '-nucleoside enzyme gene ppnN, UMP phosphohydrolase genes umpH and umpG, a ribonucleoside hydrolase 1 gene rihA and a glutamine synthetase gene glnA; an orotic acid nucleoside monophosphate decarboxylase gene pyrF and an orotic acid ribosyltransferase gene pyrE of wild type bacillus subtilis B.subtilis 168 are introduced in a heterologous manner, so that an orotate nucleoside monophosphate decarboxylase gene pyrF and an orotate ribosyltransferase gene pyrE are obtained. The safe, efficient and low-cost production of the uracil is realized.

The invention relates to the technical production field of metabolic engineering and genetic engineering, and particularly provides a genetically engineered bacterium for producing 2 '-deoxyuridine as well as a construction method and application of the genetically engineered bacterium. According to the strain, deoxyuridine kinase tdk is knocked out on an E.coli W3110 genome, and T7 RNA polymerase is integrated; an aspartate transaminase gene aspC, a glutamine synthetase gene glnA, a 5 '-deoxynucleotidase gene yfbR and a uridine monophosphate kinase gene pyrH are subjected to overexpression; the method comprises the following steps: introducing a ribonucleoside diphosphate reductase gene nrdBCA from Escherichia phage T4 in a heterologous series connection manner; the CTP synthase gene pyrG * of Corynebacterium glutamicum is introduced in a heterologous mode, and the CTP synthase gene pyrG * of Corynebacterium glutamicum is introduced in a heterologous mode. The safe, efficient and low-cost production of the 2 '-deoxyuridine is realized.

The present invention belongs to the field of microbial technology, and discloses a strain of Grifola frondosa with high extracellular polysaccharide production. The strain is named G5922, classified as Grifola frondosa, deposited on June 6, 2023, and deposited at the General Microbiological Center of China National Committee for the Preservation of Microorganisms, No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing. The present invention combines the strain selection technology of mutagenesis breeding and protoplast fusion to rearrange the genome of the Grifola frondosa strain, and screens out a strain with an extracellular polysaccharide production of 8.54 g / L, a biomass of 3.78 g / L, and a growth rate of 2.79 mm·d ‑1 The Grifola frondosa strains were increased by 67.75%, 44.27% and 8.56% respectively compared with the original strain.

The invention discloses arthrobotrys guangdongensis and application of the arthrobotrys guangdongensis in prevention and treatment of pine wood nematode diseases. The invention relates to Arthrobotrys guangdonensis 20GD2519A, which is preserved in China General Microbiological Culture Collection Center (CGMCC) on November 15, 2021, the address of the Arthrobotrys guangdonensis is No.3, Yard 1, Beichen West Road, Chaoyang District, Beijing, the postal code is 100101, and the preservation number is CGMCC No.23849. The Arthrobotrys guangdonensis 20GD2519A has the advantages that the Arthrobotrys guangdonensis 20GD2519A can be used for preparing a microbial inoculum; the invention directly aims at pathogenic nematodes, inhibits and kills nematodes, and reduces nematode density. The strain has strong sporulation ability, can maintain good hereditary stability, and still has high fatality rate to pine wood nematodes after continuous subculture. The survival rate of pine trees inoculated with the arthrobotrys guangdongensis suspension in advance can reach 72%.

The invention belongs to the technical field of plant tissue culture, and particularly relates to a method for inducing Hangzhou chrysanthemum leaf adventitious bud regeneration by using a potato extract, which sequentially comprises the following steps: cultivating Hangzhou chrysanthemum stem segments with buds into Hangzhou chrysanthemum aseptic seedlings with at least four leaves; the method comprises the following steps: taking down 3-4 leaves from the top of a sterile seedling of Hangzhou chrysanthemum, taking a leaf disc from each leaf close to a petiole by using a puncher, immediately inoculating the leaf disc onto an adventitious bud induction culture medium, and culturing in a plant tissue culture room; and obtaining Hangzhou chrysanthemum leaf adventitious buds. According to the method, the leaf disc adventitious bud induction time is short, the growth state is good, subsequent normal rooting, transplanting and breeding can be carried out, and a foundation is laid for successfully establishing a Hangzhou chrysanthemum genetic transformation system.

The invention belongs to the technical field of biology, and particularly relates to a pear somatic embryogenesis method. Comprising the following steps: collecting fruits and obtaining explants; inducing somatic embryogenesis by taking immature zygotic embryos as explants; taking the immature zygotic embryo as an explant to induce an embryogenic callus; carrying out subculture and embryogenic maintenance on the embryogenic callus; performing somatic embryogenesis on the embryogenic callus; seedling strengthening and rooting culture of regenerated plants; and acclimatizing and transplanting. The two somatic embryo induction ways taking the immature zygotic embryo and the embryonic callus as the explants are high in regeneration efficiency which can reach 100%, good in hereditary stability, good in universality and short in induction time from the explants to the regenerated plants, and are an efficient way for in-vitro rapid propagation of pears; meanwhile, the genetic transformation efficiency of pears can be improved to a certain extent, the genetic stability of transgenic plants is enhanced, and the occurrence probability of chimeras of the transgenic plants is reduced.

The invention discloses a preparation method of a purified and immortalized pilose antler stem cell line, and belongs to the technical field of biology. Comprising the following steps: S1, performing primary culture on pilose antler stem cells to obtain primary cells; s2, purifying the primary cells to obtain a purified cell strain with high multiplication capacity; and S3, carrying out natural immortalization domestication on the purified cell strain to obtain the single natural immortalization antler stem cell line. According to the method, the antler stem cells are subjected to primary culture, purification and immortalized domestication, so that the pure antler stem cell line subjected to infinite passage in a natural state can be obtained, continuous passage can be performed for more than 50 generations, the cost is effectively saved, the immortalized domestication method is realized through repeated passage, the passage is a one-passage two-passage mode, and the immortalized domestication method is simple and convenient. The repeated passage time is once every two days; meanwhile, the antler stem cell line is cloned from one cell, so that the antler stem cell line has relatively high genetic stability.

The invention belongs to the technical field of plant cultivation, provides application of bacillus H8 in cultivation of emilia sonchifolia, and particularly relates to bacillus H8, a bacillus H8 suspension with the concentration of 102-104 CFU / mL is prepared and applied to the position 2-3 cm away from the root of emilia sonchifolia as a bio-fertilizer once every 7 days, plant growth can be remarkably promoted, and the yield of emilia sonchifolia is increased. The content of medicinal components of chlorogenic acid and quercitrin in the leaves is increased; the method solves the problems of soil degradation, unstable medicinal material quality and the like caused by dependence on chemical fertilizers and pesticides in traditional cultivation, is suitable for green and efficient cultivation of emilia sonchifolia, and has the advantages of simplicity and convenience in operation, environment friendliness, low cost and the like.

The invention provides a vibrio parahaemolyticus bacteriophage RKP-VP23248 and application of the vibrio parahaemolyticus bacteriophage RKP-VP23248, the vibrio parahaemolyticus bacteriophage is preserved in the China General Microbiological Culture Collection Center on September 27, 2024, and the preservation number of the vibrio parahaemolyticus bacteriophage RKP-VP23248 is CGMCC (China General Microbiological Culture Collection Center) No. 46193. The invention further provides a bacteriophage composition, and the bacteriophage composition comprises the vibrio parahaemolyticus bacteriophage RKP-VP23248. The bacteriophage provided by the invention is separated from natural water environments such as seawater, river water and water sediment, still keeps high-level titer under continuous passage, has relatively strong genetic stability and thermal stability, and can effectively cope with disease outbreak caused by mass propagation of pathogenic bacteria when water environment conditions change. The bacteriophage has strong lysis property on the drug-resistant high-pathogenicity vibrio, the lysis rate is up to 88.51%, and the problem of vibrio drug resistance enhancement caused by antibiotic abuse can be effectively solved, so that the problems of large-scale acute death and the like of aquatic animals such as fishes, shrimps and the like caused by the high-pathogenicity vibrio are solved.

The present invention provides a 5-methyluridine producing strain and a construction method and application thereof, wherein the strain utilizes CRIPSR / Cas9 gene editing technology to produce E.coli On the UR12 genome, the artificial promoter P BBa_j23114 Replacement of uridylate kinase gene pyrH The natural promoter of umpH , umpG and no Gene, to ensure bacterial growth while enhancing the accumulation of 5-methyluridine; heterologous introduction and multiple copies of flavin-dependent thymidylate synthase gene thyX , overexpression of alkaline phosphatase gene phoA, Synergistically enhance the 5-methyluridine synthesis pathway; finally, overexpression of the serine hydroxymethyltransferase gene glyA, The supply of CH2-THF, the coenzyme of the key enzyme ThyX, is maintained, and the obtained strain has good genetic stability, high fermentation yield, and can stably produce 5-methyluridine.

The invention relates to the technical field of strain breeding, in particular to a high-yield beta-glucan Meyrowia guilliermondii strain and application thereof. According to the present invention, the high-yield beta-glucan mutant strain obtained through ARTP mutagenesis has the name of Meyerozyma guilliermondii NCUASD7, has been preserved in Guangdong Microbial Culture Collection Center on November 28, 2025, and has the preservation number of CDMCC NO: 67365; aRTP mutagenesis is high in mutation rate, good in hereditary stability and safer to operate, a large number of mutation libraries can be obtained within a short time, high-throughput primary screening and shake flask secondary screening are conducted through a 24-deep-hole plate, a yeast mutant strain with the high beta-glucan yield capacity is obtained, then subculture is conducted on the high-yield mutant strain, the hereditary stability of the strain is verified, and the high beta-glucan yield is obtained. The mutant strain not only has the characteristic of high yield of beta-glucan, but also can maintain good genetic stability.

This invention discloses a high-mannitol-producing strain of *Limosilactobacillus fermentum* and its applications, belonging to the field of biosynthesis technology. This strain, named *Limosilactobacillus fermentum* PY20 (GDMCC NO: 67749), was obtained from naturally squeezed persimmon juice. It exhibits excellent mannitol synthesis ability in fructose-based fermentation, growing and converting mannitol in a medium containing 30–60 g / L fructose with a conversion rate exceeding 69.77%. This invention overexpresses the endogenous mannitol dehydrogenase gene of PY20 in this bacterium. The resulting engineered bacterium grows and converts mannitol in a medium containing 30–60 g / L fructose with an efficiency exceeding 78.13%, representing an increase of over 11% in both mannitol yield and conversion efficiency compared to PY20, and demonstrating good genetic stability.

This invention discloses a drug-resistant Ewing sarcoma cell line, named human Ewing sarcoma cell ES 003, which was deposited at the China Center for Type Culture Collection on March 10, 2026, with accession number CCTCC NO: C202642. The cell line exhibits doxorubicin resistance; specifically, doxorubicin has an IC50 of [missing information - likely referring to an IC50 value] against the human Ewing sarcoma cell ES 003. 50 Above 5μM.

The application discloses a method for constructing a Vietnamese terminalia regenerative system, and comprises the following steps: Vietnamese terminalia stem segment test materials are obtained by grafting seedling saplings with superior branches, semi-lignified stem segments with axillary buds are used as explants, and after disinfection, the explants are inoculated into an induction culture medium to induce axillary buds, then the induced axillary buds are subjected to micro-branch cutting to carry out proliferation and rooting culture, and a large number of Vietnamese terminalia regenerative plants can be obtained. The method has the advantages of high propagation efficiency, high phenotype consistency, good genetic stability and the like, greatly shortens the production cycle of Vietnamese terminalia seedlings, reduces the variation risk of the seedlings, and provides high-quality seedlings for ecological afforestation and landscaping.

The application provides hibiscus SSR molecular markers and application thereof, and belongs to the technical field of molecular markers. The molecular markers are 15 in total, and the corresponding primer sequences are shown as SEQ ID NO. 1-30. Through detection, the polymorphism percentage of the series of markers reaches 100%, and the polymorphism information content, marker index and resolving power are excellent, and the genetic stability is strong. The molecular marker primer of the application can efficiently distinguish hibiscus germplasm materials, clearly reveals the population genetic structure and the genetic relationship, is suitable for hibiscus germplasm resource identification, genetic diversity analysis, molecular fingerprint construction and assisted breeding, and has the characteristics of strong specificity, high resolution and wide application.

The invention relates to the technical field of microorganisms, in particular to avian salmonella and application thereof. The avian salmonella is avian salmonella enteritidis, and the preservation number of the avian salmonella enteritidis is CGMCC (China General Microbiological Culture Collection Center) No.25474. The avian salmonella provided by the invention is easy to proliferate and enrich, has good growth performance, biological safety and genetic stability, and can be used as a host bacterium for proliferation of multiple strains of avian salmonella bacteriophages to realize fermentation of multiple strains of bacteriophages by one strain of host bacterium, so that a high-titer preparation of multiple salmonella bacteriophages is prepared, and the application prospect is wide. The cost is effectively reduced while the production process is simplified.

The invention provides a Shewanella algal bacterium phage (Shewanella algal bacterium phage) PKP-SH23103 and application of the Shewanella algal bacterium phage. The Shewanella algal bacterium phage is preserved in the China General Microbiological Culture Collection Center on November 29, 2023, and the preservation number is CGMCC (China General Microbiological Culture Collection Center) NO.45783. The invention further provides an application of the Shewanella algal bacterium phage PKP-SH23103. The invention further provides a bacteriophage composition, and the bacteriophage composition comprises the algal shewanella bacteriophage PKP-SH23103. The phage provided by the invention is separated from a natural water environment, still keeps high-level titer (1010PFU / mL) under continuous passage, has relatively strong genetic stability and thermal stability, has the characteristic of high permeability resistance, and can effectively cope with disease outbreak caused by mass propagation of pathogenic bacteria when water environment conditions change; the bacteriophage obtained by the invention can be used for preventing and treating diseases caused by shewanella alginate, is safe and environment-friendly, effectively replaces the use of antibiotics in a culture process, reduces the problems of water body pollution, enhanced bacterial drug resistance, drug residues and the like caused by antibiotic abuse, and has good practical application value.

This invention discloses a method for constructing and applying a marine fish muscle stem cell line. The culture medium uses basal medium as the background medium and contains LIF growth factor, EGF growth factor, and FGF2 protein derived from the leopard-gill sea bass. The final concentration of FGF2 protein from the leopard-gill sea bass is 20-30 ng / mL, 30-45 ng / mL, 45-55 ng / mL, or 55-70 ng / mL. Passaging using this medium can construct a stable, rapidly dividing, and short-passaging immortalized marine fish muscle stem cell line, capable of stable passage for more than 50 generations. This provides raw materials for exploring the function and gene regulation mechanisms of myoblasts, and provides in vitro verification experimental conditions for further research on the effects of marine fish muscle stem cells on growth rate, laying the foundation for the breeding of superior marine fish species.

The invention provides a strain for producing L-isoleucine and a construction method and application of the strain. Pck genes of the strain are knocked out, ilvBN (G20D, I21D and I22F), ilvA (V140M, F383A), lysC (T311I), hom (G378I), thrB (A20G), ppc, aspB, thrC, asd and brnFE are over-expressed, and the strain is prepared through operons Psod-ilvBN (G20D, I21D and I22F)-ilvA (V140M, F383A)-Ptuf-thrA-thrB-thrC and Psod-ppc-aspB-Ptuf

The present invention relates to a Yunnan micrococcus and its application in passivating heavy metals in livestock and poultry excrement treatment, belonging to the field of heavy metal pollution remediation. The present invention provides Yunnan micrococcus TW001, and a microbial preparation is obtained after strain activation, liquid seed preparation and liquid fermentation. The Yunnan micrococcus TW001 microbial preparation for poultry excrement passivation treatment provided by the present invention is used in soil heavy metal passivation treatment, has the advantages of fast strain growth rate, high fermentation density, strong genetic stability, good compatibility with the environment, and passivation effect on multiple heavy metals such as arsenic, cadmium and mercury, etc., is a good heavy metal biological passivator, and has good application prospects.

This invention relates to the fields of genetic engineering and microbial lipid metabolism regulation, specifically to a method for increasing the pentadecanoic acid content in *Schizochytrium*. The method includes constructing a gene knockout strain, wherein the ScAMPKα1, ScAMPKα2, or ScAMPKα3 genes are individually knocked out in *Schizochytrium* using homologous recombination technology. The amino acid sequence of the protein encoded by the ScAMPKα1 gene is shown in SEQ ID NO.1, the amino acid sequence of the protein encoded by the ScAMPKα2 gene is shown in SEQ ID NO.2, and the amino acid sequence of the protein encoded by the ScAMPKα3 gene is shown in SEQ ID NO.3. This invention is the first to knock out the adenosine monophosphate-activated protein kinase AMPKα subunit gene, effectively increasing the accumulation level of pentadecanoic acid in *Schizochytrium*.

The invention belongs to the field of microbial fermentation engineering, and particularly relates to a microbial strain with high yield of pyripyropene A and application thereof. The strain QLN-015 is preserved in the China General Microbiological Culture Collection Center on February 2, 2026, the preservation number is CGMCC No.42501, the classification name is Penicillium coprosum, and the preservation address is No.3, Yard 1, Beichen West Road, Chaoyang District, Beijing. The pyripyropene A prepared by the invention is a precursor compound of afidopyropen, belongs to a novel insecticide with a novel structure, high efficiency and low toxicity, and shows remarkable market application prospect and industrial development potential.

The invention belongs to the technical field of biomedicine, and particularly relates to a construction method and application of an anti-genome sequence and a recombinant virus, and the L delta M2-2 anti-genome sequence is shown as SEQ ID NO.2. The recombinant virus constructed by using the LdeltaM2-2 anti-genome sequence has attenuated effects of different degrees, and the recombinant virus can be used as a human or animal immune vaccine or used for preclinical experimental evaluation or can be used as a carrier carrying other antigens.

The invention discloses a porcine infectious diarrhea vaccine taking vesicular stomatitis virus as a carrier, and relates to the technical field of research and development of biological products for prevention. The porcine infectious diarrhea vaccine taking the vesicular stomatitis virus as the vector comprises a recombinant virus or an inactivated virus thereof; the recombinant virus is obtained by virus rescue by a recombinant virus vector and helper plasmids through a reverse genetic technology; the recombinant virus vector is obtained by taking VSV as a vector and replacing a G protein gene of the VSV with an S protein gene of a porcine infectious diarrhea virus. The toxicity of the vesicular stomatitis virus losing the surface envelope protein is greatly reduced, and the vesicular stomatitis virus is very suitable for being used as a virus vector. Experiments prove that the recombinant vaccine provided by the invention has good safety and effectiveness, has no neurotoxicity to mice, and can generate high serum neutralizing antibody titer at the same time. And a good neutralizing antibody titer is also generated in a piglet immune test. The invention lays a solid foundation for further research and development of PEDV vaccines, and provides a new strategy for safely and effectively preventing and controlling PEDV for the pig industry.

The invention discloses a strain capable of efficiently expressing xylanase, the strain is preserved in Guangdong Microbial Culture Collection Center on October 23, 2024, the preservation date is Pichia pastoris, and the preservation number is GDMCC NO65325. The invention further discloses a preparation method of the strain capable of efficiently expressing xylanase, and the strain is preserved in Guangdong Microbial Culture Collection Center on October 23, 2024. According to the invention, a strain with excellent genetic stability for producing xylanase is screened by means of ultraviolet light induction of the strain and component optimization and synergistic induction of a primary screening culture medium, the strain can efficiently express xylanase, and in addition, histidine and nicotinic acid are added into an enzyme production culture medium, so that the expression of xylanase is further promoted, and the yield of xylanase is improved. And the xylanase has more stable enzyme activity, so that the xylanase has better utilization value.

This invention belongs to the field of genetic engineering technology and discloses a method for synthesizing mussel adhesive protein using gene recombination technology, as well as related recombinant vectors and strains. The method uses the core functional genes of mussel byssalin 3 and 5 as templates, and constructs a recombinant expression vector pET-20b-fp-cut through artificial synthesis and overlapping PCR gene tandem. This vector is then transformed into *E. coli* BL21 to obtain the recombinant strain BL21 / pET-20b-fp-cut. A gradient-scale fermentation process is used to optimize the efficient expression of the target protein. The product is then extracted using a salting-out-acetic acid extraction combined with tyrosinase-catalyzed purification process. Finally, multi-dimensional quality testing is used to verify the indicators. This invention also provides the aforementioned recombinant expression vector and recombinant strain. This method can realize the industrial production of mussel adhesive protein, with a fermentation unit of 5.2 g / L, a dopa content of 11.2%, and a purity of 96.5%. It also exhibits low immunogenicity and meets quality standards, showing broad application prospects in biomedicine, cosmetics, and other fields.

The invention discloses an agrobacterium tumefaciens-mediated bottle gourd stable genetic transformation method. The invention provides a bottle gourd stable genetic transformation system mediated by agrobacterium tumefaciens. The system provides a key technical platform for gene function research and molecular breeding of bottle gourds. An agrobacterium tumefaciens-mediated genetic transformation method is adopted, and the principle is that a target gene is inserted into a modified T-DNA region, an agrobacterium tumefaciens infection mechanism is utilized to realize transfer and integration of an exogenous gene to a plant cell, and a transgenic plant is obtained by combining a tissue culture technology. According to the method, screening culture is not carried out, after co-culture is carried out for two days, a fluorescence microscope is used for observing the fluorescence area of a groove part of a seed notch, the agrobacterium infection efficiency is judged, screening is carried out, and detection is carried out again after seedlings are obtained, so that the time is saved by 30-40 days compared with genetic transformation of plants needing screening culture; the method has the advantages of good hereditary stability and simple operation.

This invention discloses a Hunan pig SNP molecular marker and its application. The Hunan pig SNP molecular marker of this invention includes at least one of 64,055 SNP molecular markers. The genomic positions of these 64,055 SNP molecular markers were determined by sequence alignment based on the pig reference genome Sscrofa11.1. The Hunan pig SNP molecular markers of this invention are derived from the diverse local pig breeds of Hunan, exhibiting broad coverage, good site specificity, strong universality, and high accuracy. They can effectively distinguish local pig breeds from different origins and provide important technical support for the molecular breeding application of Hunan pigs.

This invention provides a uracil-producing strain, its construction method, and its application. The strain utilizes CRIPSR / Cas9 gene editing technology to produce... E. coli On the UR14 genome, firstly by knocking out psuG Gene, black Gene, rutA Genes and upp Genes were introduced to block the breakdown of uracil; then, the uridine phosphorylase gene was overexpressed. udp and pyrimidine-5'-nucleotide nuclease gene ppnN This synergistically enhances the synthesis and accumulation of uracil; finally, overexpression of the phosphoribosyltransferase gene... pgm, Further enhancing the conversion of the byproduct 1-phosphate ribose into the precursor 5-phosphate ribose-1-pyrophosphate improves carbon utilization. The resulting strain exhibits good genetic stability and high fermentation yield, enabling stable production of uracil and demonstrating broad application prospects.