20results about How to "High genetic stability" patented technology

The invention discloses a methionine nutrition selection marker saccharomyces cerevisiae vector as well as a construction method and application thereof, and relates to the field of genetic engineering and synthetic biology, a methionine MET17 promoter and a coding region sequence are obtained through gene artificial synthesis, a leucine promoter and a coding region sequence in a pAG415-GAL1-ccdB vector are replaced by homologous recombination, and thus the methionine nutrition selection marker saccharomyces cerevisiae vector is obtained. A pAG415-GAL1-MET-ccdB expression vector is constructed and obtained, a ccdB gene is replaced by a green fluorescent protein reporter gene GFP sequence to be integrated into the pAG415-GAL1-MET-ccdB vector, a pAG415-GAL1-GFP-MET vector is constructed and obtained, the obtained expression vector can be used for screening exogenous gene transformants in a methionine deficient culture medium, meanwhile, green fluorescent GFP tag protein carried in the vector has the advantages that the expression vector can be applied to screening of the methionine deficient culture medium, and the expression efficiency of the methionine deficient culture medium is improved. And more application space is provided for carrying out subcellular localization, immunoprecipitation and other downstream inheritance and molecular biology analysis on a target gene in wine brewing cells.

A method for cryopreservation and recovery of larch olgensis embryogenic callus relates to the biotechnology field of forest breeding, and comprises the following steps: selecting an embryogenic cell line induced by immature seeds of larch olgensis, and subculturing the embryogenic cell line for 3-5 days to obtain embryogenic callus as a standby material; carrying out gradient pre-culture at the molar concentrations of 0.2 mol / L and 0.4 mol / L in sequence, wherein the culture is carried out at each concentration for 20-48 hours; after pre-culture, putting the embryogenic callus into a freezing tube, and adding a sterile freezing protection solution; and putting the freezing pipe into a programmed cooling box for programmed cooling, and immediately putting the freezing pipe into a liquid nitrogen biological container for storage. The method disclosed by the invention is simple and convenient in operation steps and obvious in effect, the subculture age limit of the larix olgensis embryonic callus is slowed down, and relatively good cell viability and development potential in the initial stage of induction are maintained, so that the occurrence of variation is effectively reduced, manpower and material resources consumed by long-term subculture materials can be saved, and the production cost is reduced. The method is of great significance to theoretical and practical application research of larix olgensis.

The invention relates to the technical field of plant genetic engineering and agricultural planting, in particular to application and method of an SlGRXC9 gene in improvement of tomato cadmium stress tolerance, the number of the SlGRXC9 gene is Solyc07g053550, the core coding region sequence of the SlGRXC9 gene is SEQ ID NO.1, and the coded protein sequence is SEQ ID NO.2; through construction of SlGRXC9 overexpression transgenic tomatoes, heterologous expression of protein encoded by the gene in plant cells or use of the gene as a molecular breeding target, the system activity of antioxidant enzymes such as SOD, POD, CAT and GR in tomatoes can be enhanced, cadmium stress induced H2O2 and O2-accumulation and membrane lipid peroxidation damage are reduced, and absorption and accumulation of the tomatoes to cadmium ions are not affected; according to the method, the growth ability and physiological stability of the tomatoes under cadmium stress are remarkably improved, the risk of nutrient imbalance is avoided, the hereditary stability is high, and the method can be widely applied to safe production of the tomatoes in cadmium-contaminated soil and creation of stress-tolerant germplasm.

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 present invention relates to a recombinant halomonas comprising an exogenously introduced xylose metabolic pathway and a metabolic pathway for synthesizing a copolymer of hydroxybutyric acid (HB) and lactic acid (LA), whereby the copolymer can be synthesized from a xylose-containing cellulose hydrolysate, the LA ratio can be controlled by further promoter modification and process optimization, and industrial enlargement is facilitated.

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 invention discloses a method for medium-short-term preservation and growth recovery of pinellia ternate. The method comprises the steps of material culture and disinfection, second-generation test-tube plantlet and second-generation tuber culture, medium-short-term preservation of third-generation tubers, growth recovery and propagation expansion. The Pinellia ternate medium-short term preservation technology can be applied to Pinellia ternate germplasm resource preservation, compared with field preservation, the Pinellia ternate medium-short term preservation technology can greatly save manpower and material resources, is easy and convenient to operate and not prone to being attacked by natural disasters, has the advantages of being small in occupied space, low in preservation cost, long in preservation time, stable in heredity and the like, and Pinellia ternate germplasm can be preserved in a medium-short term.

The invention relates to the technical field of microorganisms, in particular to streptomyces noursei for producing multi-fungal element B and a method for increasing the yield of the multi-fungal element B. According to the streptomyces noursei for producing the multi-fungal element B, negative regulation of the multi-fungal element B is relieved or negative regulation is relieved and positive regulation is enhanced through a combined genetic engineering strategy, so that the yield of the multi-fungal element B is remarkably increased, and byproducts are remarkably reduced; in addition, the strain also has relatively high genetic stability. The method for increasing the yield of the polymycotin B of the streptomyces nosella provides an effective transformation target for construction of a polymycotin B production strain, and has a relatively good application prospect in the production of the polymycotin B. The invention further provides a method for improving the yield of the polymycotin B of the streptomyces nosella.

The application discloses a recombinant Yarrowia lipolytica engineering bacterium for synthesizing borneol from scratch, a construction method and application thereof. The recombinant Yarrowia lipolytica engineering bacterium is constructed by integrating a multi-gene tandem expression cassette containing an orange-flower base pyrophosphate synthase coding gene, a borneol base pyrophosphate synthase coding gene and a hydrolytic enzyme coding gene into a genome H1 site of Yarrowia lipolytica ATCC 201249 by CRISPR-Cas9 gene editing technology. The application first constructs the recombinant Yarrowia lipolytica engineering bacterium for synthesizing borneol from scratch by using a simple carbon source. The engineering bacterium does not need to add any terpene precursor externally, and only uses simple carbon sources such as glucose and sucrose as substrates. The borneol is synthesized from acetyl coenzyme A, a primary metabolic starting material, by a complete MVA pathway and an artificially constructed borneol-specific synthesis pathway, so that the whole biosynthesis of borneol is realized, and the limitation of the prior art on external precursors is broken through.

The application relates to the fields of molecular biological technology and livestock breeding technology, in particular to a SNP molecular marker related to a total number of piglets character, a detection primer pair, a kit and application of the SNP molecular marker in a method for breeding large white pigs with high number of piglets, the SNP molecular marker is located on a pig 9th chromosome, a physical position is located at chr9:65633096 of a genome version Ensembl Sscrofa11.1, a polymorphism site is C or A, and the C allele is positively correlated with the high number of piglets character; through genotype discrimination, selection breeding can be carried out: an individual with a genotype of C / C is preferentially selected or reserved as a breeding pig, because the total number of piglets has the highest potential, so that the genetic improvement speed of a population breeding performance is accelerated.

The invention discloses a cryopreservation method of a sterile stem tip of terpinen-4 alcohol type melaleuca alternifolia. Comprising the following steps: (1) obtaining sterile stem tips; (2) pretreatment of sterile stem tips; (3) loading the sterile stem tips; (4) dehydrating the sterile stem tips; (5) cryopreservation; (6) thawing in a water bath; (7) unloading; and (8) carrying out recovery culture and rooting culture. According to the method disclosed by the invention, the minimization of genetic variation is ensured, the time and labor cost of explant disinfection, primary culture and the like are greatly saved, the problem of'senescence 'phenomenon caused by long-term mass subculture in the current production is avoided, and the long-term preservation of the sterile stem tips is realized for the first time.

The invention relates to the technical field of biology, and discloses a recombinant plasmid for vaccine preparation, a preparation method of the recombinant plasmid and an attenuated live hand-foot-and-mouth disease vaccine started by DNA (Deoxyribose Nucleic Acid). The recombinant plasmid provided by the invention comprises an expression vector, an insertion element and a virus genome sequence, the expression vector is selected from a eukaryotic expression vector; the insertion element is selected from a eukaryotic promoter and an optional element capable of improving the expression of the recombinant plasmid; the virus genome sequence is selected from an enterovirus 71 type genome sequence. The recombinant plasmid has the characteristics of high genetic stability, high safety, low requirements on transportation and preservation conditions and the like, experiments prove that the recombinant plasmid can rescue in RD cells and normally start transcription and express virus proteins, shows good safety and immune protection effects in animal experiments, and has the potential of preparing DNA-started attenuated live hand-foot-and-mouth disease vaccines.

The invention discloses an efficient implementation method for wheat breeding, and relates to the technical field of crop genetic breeding, and the method comprises the following steps: S1, setting a parent screening index system based on a target breeding demand, analyzing and calibrating indexes through big data, and screening parent materials conforming to the index system, hybridizing by using a flowering phase regulation and control technology and artificial pollination to obtain F1-generation seeds; and S2, constructing a filial generation stepped cultivation system, and carrying out expanding propagation from F1-generation mixed germplasm to F4-generation strain planting. According to the efficient implementation method for wheat breeding, by comprehensively applying technical means such as big data analysis, molecular marker-assisted selection and a stepped cultivation system, the efficiency and accuracy of wheat breeding are remarkably improved, parent screening indexes are calibrated by means of big data, the suitability of genetic differences of parent materials is ensured, and the method is suitable for popularization and application. A foundation is laid for polymerization of excellent characters, the breeding period is effectively shortened, and the completeness of standard execution and data recording is ensured.

This invention discloses a method for improving the stability of arginine production capacity in bacterial strains, comprising the following steps: using *Corynebacterium glutamicum* producing L-arginine as the base strain, inactivating or attenuating the gene NCgl2644 / cg3035 in the genome. The method of this invention can effectively improve the genetic stability of L-arginine-producing strains.

The invention discloses a molecular marker of a rice cold-resistant gene and application thereof, and relates to the field of rice breeding. The molecular marker is located in a promoter region (-307-364) of a rice seedling stage cold-resistant gene LTSS9a (LOCOs09g21710). The invention provides a primer for detecting the molecular marker of the rice seedling stage cold-resistant gene and a kit comprising the primer. The invention also provides application of the molecular marker in identifying the cold resistance of the rice in the seedling stage and a method for identifying the cold resistance of the rice in the seedling stage. The molecular marker provided by the invention has high accuracy and genetic stability, is stable in performance under different generations and environmental conditions, and is not easy to mutate; the method is simple and convenient to operate and visual in result, and can be used for rapidly identifying the rice seedling-stage cold-resistant variety, so that a powerful tool is provided for research on rice cold resistance, and the problem of rice yield reduction caused by low-temperature cold damage is effectively solved.

The invention discloses an artificial propagation-hybridization combined breeding method for meat goats, and relates to the technical field of animal breeding, and the method comprises the following specific steps: selecting a goat variety with good meat performance as a female parent, and selecting a goat variety with large physique as a male parent; the method comprises the following steps: performing in-vitro culture on female parent oocytes to obtain mature oocytes containing first polar bodies, screening high-activity sperms by adopting a temperature gradient screening device, performing in-vitro fertilization on the mature oocytes and the high-activity sperms, culturing the mature oocytes and the high-activity sperms to a hybrid blastocyst form, transplanting the mature oocytes and the high-activity sperms into a female body, and developing the mature oocytes and the high-activity sperms until the mature oocytes and the high-activity sperms are naturally delivered to obtain a hybrid F1 generation; backcrossing the F1 generation with the female parent, backcrossing the backcrossed offspring with the female parent for multiple times until a breeding target is met, and carrying out cross-crossing fixation to obtain a new variety / line with excellent characters; the sperm screening efficiency is improved by utilizing the screening device, and the quality of the screened sperms is ensured, so that the fertilization efficiency and the embryo survival rate are remarkably improved.

The present invention provides a recombinant Yarrowia lipolytica of high-yield tetraacetyl phytosphingosine and its application, belonging to the field of bioengineering. The recombinant Yarrowia lipolytica of high-yield tetraacetyl phytosphingosine constructed by the present invention is named as Yarrowia lipolytica (Yarrowia lipolytica) XJ TAPS strain, and the construction method of the recombinant Yarrowia lipolytica of high-yield tetraacetyl phytosphingosine, including Wickerhamomyces ciferrii (Wickerhamomyces ciferrii) phytosphingosine acetyltransferase enzyme gene expression cassette, phosphorylase gene knockout cassette, serine palmitoyltransferase gene expression cassette, 3 keto dihydrosphingosine reductase gene expression cassette, C 4 hydroxylase gene expression cassette, integrated in the Yarrowia lipolytica genome by homologous recombination. The construction method of the recombinant Yarrowia lipolytica of the present invention is simple to operate, and tetraacetyl phytosphingosine can be efficiently synthesized.

The invention belongs to the field of bioengineering, and provides recombinant yarrowia lipolytica with high yield of beta-caryophyllene as well as a construction method and application of the recombinant yarrowia lipolytica. The recombinant yarrowia lipolytica with high yield of beta-caryophyllene is named as a yarrowia lipolytica XJ-CAP strain, the recombinant yarrowia lipolytica XJ-CAP strain can be used for producing 6.27 g / L of beta-caryophyllene after being subjected to fed-batch fermentation, and the efficient synthesis of a plant source natural product beta-caryophyllene in the yarrowia lipolytica is realized. The construction method of the recombinant yarrowia lipolytica is easy to operate, and efficient synthesis of beta-caryophyllene can be achieved.

The invention discloses a method for directionally screening a teriparatide high-yield strain by using antibiotics, an escherichia coli production strain capable of highly expressing teriparatide is obtained through two rounds of screening of antibiotics with different concentrations, and the stability of plasmids in a strain passage process is still effectively maintained under the condition that no antibiotics are added. According to the directional screening method, the screening efficiency of the teriparatide high-yield strain can be effectively improved, and the technical difficulty that antibiotics need to be added in the production process of biological drugs is overcome.

The invention discloses a seed production method and application of a terminal paternal line for producing tile grey chickens with crested characters, and belongs to the technical field of poultry genetic breeding and propagation. According to the method, the grey character of the tile grey chicken and the appearance character of the crested head of the Taihe black-bone chicken are successfully combined for the first time, high-quality gene resources of the tile grey chicken and the Taihe black-bone chicken are effectively integrated, the genetic advantages of local varieties are exerted, and a feasible path is provided for innovative utilization of local poultry varieties; by utilizing a molecular marker-assisted breeding mode, the genetic stability of target traits is directionally enhanced, the breeding period is shortened, and the breeding efficiency is remarkably improved; a new poultry strain with white feather is used as a terminal paternal line to be hybridized with other colored feather lines, so that commercial generation tile grey chickens with gray feather can be produced, and the difficulty of breeding the same commercial line by using commercial generation broiler chickens and the difficulty and cost of reverse imitation are greatly increased. The method has wide application prospects in the aspects of characteristic poultry breeding, ornamental poultry breeding and high-quality poultry breeding.