399 results about "Gene engineering" patented technology

The invention belongs to the technical field of gene engineering, and particularly relates to mutants of murA and ygeA genes and application of the mutants in production of human milk oligosaccharides. According to the invention, murA and ygeA genes of escherichia coli are edited by using a gene editing technology, so that a genetic engineering chassis strain carrying murA and ygeA mutants is obtained. Further, the chassis strain is applied to the construction process of production strains of LNT II, LNnT, LNT, 3 '-SL and 6'-SL, and the yield of the LNT II, the LNnT, the LNT, the 3 '-SL and the 6'-SL is further increased.

The invention belongs to the technical field of gene engineering, and particularly relates to applications of OpgG and a mutant thereof in production of human milk oligosaccharide. The invention provides a dextran biosynthetic protein OpgG mutant, and the OpgG mutant is obtained by mutating the 443rd amino acid from threonine to proline on the basis of a wild type dextran biosynthetic protein OpgG as shown in SEQ ID NO.1. The invention also provides a preparation method of the dextran biosynthetic protein OpgG mutant. An opgG gene and / or a T443P mutant coding gene are / is introduced into a human milk oligosaccharide production strain and applied to production of human milk oligosaccharide, and the yield of 3 '-SL, 6'-SL, LNT II, LNnT and LNT is greatly increased.

The invention discloses recombinant escherichia coli for inactivating or weakly expressing SPMB family inner membrane protein and application of the recombinant escherichia coli in production of L-amino acid. The invention relates to the technical field of gene engineering, and provides recombinant escherichia coli, and SPMB family inner membrane protein in the recombinant escherichia coli is inactivated or reduced in activity. According to the invention, the yjiG gene (coding gene of SPMB family inner membrane protein) in escherichia coli is knocked out, so that the expression intensity of the yjiG gene is weakened, and fermentation production strains of L-threonine, L-tryptophan, L-arginine and L-valine, which are more cost-effective, can be produced. The invention has important significance for improving the fermentation yield of bacteria, namely L-threonine, L-tryptophan, L-arginine and L-valine.

The invention relates to the technical field of gene engineering, in particular to tyrosine sulfated recombinant hirudin as well as a preparation method and application thereof. In order to improve the safety and synthesis yield of the recombinant hirudin and reduce the production cost, the tyrosine sulfated recombinant hirudin is successfully prepared through an algae expression vector.

The invention discloses application of TaMYB-7B and a coding gene thereof in regulation and control of wheat tiller number, and belongs to the technical field of gene engineering. The invention finds for the first time that the TaMYB-7B gene is a novel gene capable of simultaneously regulating and controlling the tillering number and grain traits of wheat. The TaMYB-7B gene is interfered, knocked out, silenced or mutated, so that the tiller number of a plant can be increased, the grain length is increased, the hundred-grain weight is increased, finally, the yield of a single wheat plant can be increased, and a foundation is laid for wheat yield improvement and character improvement.

The invention relates to the technical field of gene engineering and enzyme engineering, in particular to an esterase mutant for degrading ethyl carbamate and application of the esterase mutant, the esterase mutant has an amino acid sequence as shown in SEQ ID NO.1 which is subjected to site mutation, the site mutation is any one or more site combined mutation of V129S, V161L or I229M, and the amino acid sequence of the esterase mutant is shown in SEQ ID NO.1. The V129S site mutation is that valine at the 129 site is mutated into serine, the V161L site mutation is that valine at the 161 site is mutated into leucine, and the I229M site mutation is that isoleucine at the 229 site is mutated into methionine. Mutation is carried out through a site-directed mutagenesis method, so that the amino acid sequence of the esterase mutant V129S / I229M is changed, the autoplasmic structure and function of eggs are changed, the esterase mutant V129S / I229M with the ethyl carbamate hydrolysis rate increased to 6.55 times in an acid environment is finally obtained, and the esterase mutant V129S / I229M has high industrial application value.

The invention discloses a gene for enhancing gray mold resistance of strawberry fruits and application of the gene, and belongs to the technical field of gene engineering. The nucleotide sequence of the gene disclosed by the invention is as shown in SEQ ID NO.7. The invention provides the gene for enhancing the gray mold resistance of the strawberry fruit, the gene is introduced into the strawberry body to realize stable overexpression, the gray mold resistance of the strawberry fruit is obviously enhanced, the gene has no influence on the quality of the strawberry fruit, and a new gene resource and a theoretical support are provided for breeding of strawberry varieties with high gray mold resistance.

The invention provides an application of citrus flavonoid glycosyl transferase CitUGT73A78 in biosynthesis of flavonoid glycoside, and belongs to the technical field of gene engineering and enzyme engineering. According to the present invention, the function of the CitUGT73A78 gene is cloned and verified, and the in vitro enzyme activity test proves that the CitUGT73A78 recombinant protein has broad-spectrum catalytic activity, and can efficiently catalyze the glucosylation of a variety of flavonoids so as to generate the corresponding flavonoids 7-O-glucoside and / or flavonoids 4 '-O-glucoside. An in-vivo transient overexpression test also proves that the CitUGT73A78 gene has a promoting effect on the accumulation of the content of the citrus flavonoid 7-O-glucoside. The invention not only provides a preparation method for the production of flavonoid 4 '-O-glucoside, has the advantages of simplicity, convenience, high catalytic efficiency, low cost, greenness and safety, but also can be applied to nutritional quality improvement and germplasm innovation of citrus fruits.

The invention relates to the field of gene engineering and enzyme engineering, in particular to an acid protease mutant with improved stability and a product and application thereof. In particular to an acid protease mutant IAIC6M with improved heat resistance and a coding gene thereof. A mutant IAIC6M is obtained by performing D86I, G168A, Q185I and Q339C mutation on an iaic6 sequence as shown in an amino acid sequence SEQ ID NO.1, and compared with IAIC, the mutant can still maintain more than 60% of enzyme activity at 75 DEG C for 3 minutes, while the IAIC can only maintain 20% of enzyme activity at 75 DEG C for 3 minutes; the acid protease mutant IAIC6M disclosed by the invention has the following properties: the optimum pH is 3.0, and the acid protease mutant IAIC6M still has higher enzyme activity under a high-temperature condition and has wide application value in the aspects of industry, food, medicine and the like.

The invention belongs to the technical field of gene engineering, and particularly relates to a mutant of polymyxin efflux transporter and application of the mutant. The mutant is a PmxD transporter mutant, the amino acid sequence of the PmxD transporter mutant is shown as SEQ ID NO: 1, and compared with wild type PmxD, the polymyxin transport capacity of the PmxD transporter mutant (T38W) is improved by 450.46%; and the total discharge amount of polymyxin is increased by 85.72%. Meanwhile, the mutant can significantly improve the growth ability of the strain on a plate containing 250 [mu] g / mL of polymyxin B, namely significantly improve the autoresistance of paenibacillus polymyxa to polymyxin.

The invention belongs to the technical field of biological catalysis, and particularly relates to four R-transaminase and application thereof in asymmetric synthesis of chiral N-heterocyclic amine active drug intermediates. The amino acid sequences of the R-transaminase disclosed by the invention are shown as SEQ ID NO. 1, SEQ ID NO. 3, SEQ ID NO. 5 and SEQ ID NO. 7. Four kinds of R-transaminase are obtained through gene mining and screening, and recombinant escherichia coli is constructed and expressed by adopting a gene engineering technology. The obtained R-transaminase can be used for synthesizing chiral drug intermediates (R)-3-amino-N-Boc-pyrrole (1b) and (R)-3-amino-N-Boc-piperidine (2b) with high stereoselectivity, and has a good application prospect.

The invention discloses SGNH family esterase SH2 screened in a Tibetan hot spring environment and application of the SGNH family esterase SH2, and belongs to the technical field of gene engineering. According to the invention, a new SGNH family esterase gene is screened from a Tibetan hot spring environment, and recombinant expression is realized in escherichia coli. The recombinant esterase prepared by the invention has the characteristics of thermal stability, salt resistance, organic solvent resistance and the like, can be safely applied to tooth whitening or skin exfoliating products, avoids damage of traditional chemical bleaching agents to enamel, and has important application value in the fields of daily chemicals, medicines, food processing and the like.

The invention belongs to the technical field of gene engineering, and discloses an acid protease mutant as well as a coding gene and application thereof. Compared with parent acid protease with an amino acid sequence as shown in SEQ ID NO: 1, the acid protease mutant comprises mutant S60V; the acid protease mutant has at least 98% and less than 100% sequence identity with the parent acid protease. Compared with parent acid protease, the thermal stability of the acid protease mutant provided by the invention is remarkably improved, and the enzyme activity retention rate is greater than 63% after the acid protease mutant is treated at 75 DEG C for 3 minutes and can reach 90.6% to the maximum. The acid protease mutant provided by the invention is beneficial to the application of protease in industrial production.

The invention discloses an endoglucanase mutant with acid resistance and application of the endoglucanase mutant, and belongs to the technical field of gene engineering and enzyme engineering. According to the present invention, PCR amplification is adopted to carry out site-specific saturation mutation on endoglucanase tCel5A, and n variants S45D / T55D / T59D and T88H / W255H are constructed and screened; the enzyme activity determination result shows that the enzyme activities of the endoglucanase variants S45D / T55D / T59D and T88H / W255H under the acidic condition are 8828.7 U / mg prot and 5206.7 U / mg prot respectively, and the degradation rates of the endoglucanase variants S45D / T55D / T59D and T88H / W255H to vinasse under the solid-to-liquid ratio of 1: 30 are 53.53% and 50.47% respectively, so that the endoglucanase variants S45D / T55D / T59D and T88H / W255H have high acid resistance and plant cellulose degradation capacity and have great application value in the aspects of agriculture and brewing.

The invention discloses a high-activity endoglucanase expression optimized strain and application thereof, and belongs to the technical field of gene engineering and enzyme engineering. According to the present invention, PCR amplification and homologous recombination are adopted to carry out strain construction and expression optimization on endoglucanase tCel5A, and screening is performed to obtain the sAOX1-pSUC2-tCel5A strain; an enzymatic property research finds that the recombinant pPICZA-tCel5A has the optimal temperature of 50 DEG C and the optimal reaction pH of 4.5, and has good thermal stability and low pH stability. The enzyme activity determination result shows that the enzyme activity of the sAOX1-pSUC2-tCel5A after expression optimization is 4632.81 U / mg prot, which indicates that the tCel5A after expression optimization can efficiently degrade cellulose, and has a great application prospect in the fields of agriculture, brewing industry and the like.

The invention discloses an inositol dehydrogenase mutant as well as a preparation method and application thereof, and belongs to the technical field of gene engineering. The inositol dehydrogenase mutant is obtained by mutating an amino acid sequence as shown in SEQ ID NO.1, and the mutation is that the 247th amino acid is mutated from K to R, or the 266th amino acid is mutated from E to D. The inositol dehydrogenase mutant disclosed by the invention has higher enzyme activity, and when the inositol dehydrogenase mutant and ketoisomerase are catalyzed to synthesize D-chiral inositol by taking myo-inositol and NADP < + > as substrates, the reaction is more inclined to a forward reaction, so that the yield of the D-chiral inositol and the conversion rate of the substrate myo-inositol are further improved.

The invention discloses application of an upstream non-coding region uORF of an ABA2 gene in regulation and control of crop agronomic traits, in particular to application in improvement of pre-harvest sprouting resistance of rice and cultivation of pre-harvest sprouting resistant rice strains, and belongs to the technical field of gene engineering. The invention finds that uORF in the upstream non-coding region of the crop ABA2 gene can inhibit the translation level of downstream ABA2 protein on the basis of not influencing the transcriptional level, so that the uORF can participate in regulation and control of crop agronomic traits in which ABA participates in regulation and control, such as improvement of pre-harvest sprouting resistance of rice. Therefore, the upstream non-coding region uORF of the crop ABA2 gene can be modified through a hybridization or gene editing technology, so that the purpose of regulating and controlling crop agronomic traits in which ABA participates in regulation and control is achieved, for example, site-directed mutagenesis is performed on an initiation codon of the upstream non-coding region uORF of the rice ABA2 gene, and the pre-harvest sprouting resistance of rice is improved.

The invention belongs to the technical field of gene engineering, and provides shRNA (short hairpin Ribonucleic Acid) for knocking down BST2 gene expression, a recombinant vector and application of the shRNA, and the shRNA comprises Bst2-shRNA-1 or Bst2-shRNA-2; the Bst2-shRNA-1 comprises a positive-sense strand of which the nucleotide sequence is as shown in SEQ ID NO. 1 and an antisense strand of which the nucleotide sequence is as shown in SEQ ID NO. 2; the Bst2-shRNA-2 comprises a positive-sense strand of which the nucleotide sequence is as shown in SEQ ID NO. 3 and an antisense strand of which the nucleotide sequence is as shown in SEQ ID NO. 4. The shRNA disclosed by the invention plays an important role in inhibiting non-infectious aseptic inflammation, can effectively delay the ALS attack time and protect motor neurons, has a very good application prospect, and also provides a direction and a technical basis for searching, researching and developing new therapeutic drugs for treating ALS.

The invention belongs to the technical field of gene engineering, and particularly relates to an LgtA mutant and application thereof in production of human milk oligosaccharide. According to the present invention, a gene editing technology is adopted to mutate a key enzyme beta-1, 3-N-acetyl glucosamine transferase for synthesizing an intermediate product LNT II so as to obtain a mutant having at least one of R13H mutation, L24M mutation and R205C mutation on the basis of a wild type LgtA represented by SEQ ID NO.1; further, the mutant is applied to the construction process of production plasmids of the LNTII, the LNnT and the LNT, and the yield of the LNTII, the LNnT and the LNT is further increased.

The invention relates to the technical field of gene engineering, in particular to a glycosyl transferase mutant, a sucrose synthase mutant and application of the glycosyl transferase mutant and the sucrose synthase mutant in synthesis of rebaudioside M. Wild glycosyl transferase and wild sucrose synthase are modified according to a directed evolution theory to obtain a glycosyl transferase mutant and a sucrose synthase mutant. Rebaudioside M is obtained by catalyzing rebaudioside D through a glycosyl transferase mutant and sucrose synthase mutant co-expression strain, and the catalytic efficiency is superior to that of a wild enzyme. The rebaudioside M is obtained by completely catalyzing 110 g / L of rebaudioside D through the co-expression strain of the glycosyl transferase mutant and the sucrose synthase mutant, only 20 h is needed, the conversion rate reaches 97%, the yield of the rebaudioside M reaches 122 g / L, the raw material conversion rate and the yield of the rebaudioside M are effectively increased, and meanwhile the synthesis time is shortened.

The invention belongs to the technical field of gene engineering, and particularly relates to a vitamin D3 hydroxylase mutant and application thereof in preparation of 25-hydroxyvitamin D3. According to the invention, the vitamin D3 hydroxylase Vdh-K1 is subjected to directed mutation, so that the vitamin D3 hydroxylase mutants VM, MV and VMV with higher activity are obtained. Compared with parent vitamin D3 hydroxylase Vdh-k1, the activities of the mutants VM, MV and VMV are respectively improved by 2.4 times, 1.6 times and 2.2 times, and when the mutant is used for preparing the 25-hydroxyvitamin D3, the conversion rate and the conversion rate of converting VD3 into the 25-hydroxyvitamin D3 are obviously improved.

The invention discloses an application of HuR and a coding gene thereof in preparation of a medicine for treating senile deafness, belongs to the technical field of gene engineering, and provides an application of the HuR coding gene in preparation of a medicine for preventing or treating senile deafness, and the nucleotide sequence of the HuR coding gene is shown as SEQ ID NO.1. According to the application of the HuR in preparation of the medicine for preventing or treating the senile deafness, the amino acid sequence of the HuR is shown as SEQ ID NO.2. The invention discovers the application of HuR in preventing or treating age-related hearing loss for the first time. The HuR coding gene is packaged by an adeno-associated virus (AAV) and then is injected into the inner ear through a round window, so that the HuR coding gene is widely overexpressed in various cells of the inner ear, the occurrence of the senile deafness can be resisted or delayed, and a theoretical basis is provided for clinically preventing and improving the hearing loss of a patient with the senile deafness.

The invention belongs to the field of gene engineering. Specifically, the invention relates to a base editing system for optimizing engineered T cells and application of the base editing system. The present invention provides a base editing system for modifying engineered T cells, comprising: i) a base editing polypeptide, and / or an expression construct encoding a base editing polypeptide, the base editing polypeptide comprising a DNA binding protein and at least one deaminase domain; ii) a guide RNA, and / or an expression construct encoding a guide RNA, the guide RNA targeting any one of TRAC, TGFBR2, PD-1, CISH, and / or FAS genomic regions, or any combination thereof. The base editing system disclosed by the invention has high efficiency of a gene editing tool and low Indel frequency, can realize accurate editing of T cell function enhancement related genes, and has important value for expanding application scenes of a base editing technology in the fields of preparation of safe and efficient engineered T cells and the like.

The invention provides application of an OsRTD1 gene in improving drought resistance of rice, and belongs to the technical field of gene engineering and rice drought resistance. The nucleotide sequence of the OsRTD1 gene is as shown in SEQ ID NO: 1, and the amino acid sequence of the protein coded by the OsRTD1 gene is as shown in SEQ ID NO: 2. Compared with a wild plant, the survival rate of the OsRTD1 gene mutant plant osrtd1 under the drought stress of PEG induction and soil dehydration is remarkably improved, the survival rate of the OsRTD1 gene overexpression plant OsRTD1-OE under the drought stress is remarkably reduced, the relative water loss rate of dehydrated leaves is higher, and the survival rate of the OsRTD1 gene overexpression plant OsRTD1-OE under the drought stress is remarkably reduced. The total grain number, the solid grain number, the total grain weight and the like of the OsRTD1 gene mutant plant osrtd1 under the stress of field drought are obviously higher than those of a wild plant. The OsRTD1 gene is a potential candidate new target for regulating and controlling a rice drought resistance mechanism.

The invention discloses an electron transport chain subunit acCYTB with low-temperature activity and application of the electron transport chain subunit acCYTB, and relates to the technical field of gene engineering and the technical field of enzyme engineering. The electron transport chain subunit provided by the invention is any one of the following (1), (2) and (3): (1) acCYTB protein, the amino acid sequence of which is as shown in SEQ ID NO.2; (2) carrying out substitution, deletion and / or addition of one or more amino acid residues on the amino acid sequence of the acCYTB protein to obtain a protein which has more than 90% of identity with the acCYTB protein and has the same biological function with the acCYTB protein; and (3) a fusion protein obtained by connecting a protein tag to the N terminal or / and C terminal of (1) or (2). The electron transport chain subunit can improve the low-temperature resistance of organisms, so that the electron transport chain subunit is applied to low-temperature character improvement and industrial production.

The invention provides application of an Aars1 gene in regulation and control of expression of an exogenous recombinant protein, and belongs to the technical fields of recombinant protein expression, gene engineering and cell culture. According to the CHO cell line, shRNA fragments of targeted Aars1 genes are inserted and integrated into piggyBac transposon vectors, a recombinant vector for inhibiting / silencing the Aars1 genes and an Aars1 low-expression CHO cell line are constructed, the CHO cell line can remarkably reduce the mRNA expression level of the Aars1, meanwhile, the CHO cell line serves as a host cell for recombinant protein expression, and compared with a wild type CHO cell line, the CHO cell line has the advantages that the expression level of the CHO cell line is greatly improved; the recombinant adalimumab can improve the expression quantity of recombinant protein (the expression quantity of the recombinant adalimumab is increased by 3.1 + / -0.18 times), effectively overcomes the problem of low expression level of the existing CHO cell expression system, and can be used for further constructing a recombinant protein expression system or kit.

The invention relates to the technical field of gene engineering, and particularly discloses streptococcus agalactiae WC1535 delta cps delta hylB as well as a construction method and application thereof. The streptococcus agalactiae WC1535 [delta] cps [delta] hylB is preserved in Guangdong Microbial Culture Collection Center on February 28, 2025, the preservation number is GDMCC NO.65955, and the address of the preservation unit is the 5th building, No. 59 building, No. 100 Courtyard, Xianlie Middle Road, Guangzhou. Compared with a traditional single-fragment deletion method, the toxicity of the deletion strain is not prone to being enhanced, and compared with a traditional injection immune vaccine, the gene deletion strain is used for preparing the tilapia oral live vaccine, the immune operation is simpler, and more labor cost is saved.

The invention discloses a carbonyl reductase mutant as well as a preparation method and application thereof, and belongs to the technical field of gene engineering and enzyme engineering. According to the invention, wild-type carbonyl reductase of Stachybotrys chlorohallonata is taken as an evolution template, and a plurality of mutants with catalytic activity, thermal stability and substrate spectrum obviously superior to those of the wild-type carbonyl reductase are obtained through an enzyme engineering technology. The carbonyl reductase mutant can catalyze various prochiral carbonyl compounds to obtain chiral alcohol with high stereoselectivity (ee is greater than 99%). The carbonyl reductase mutant provides a novel, efficient and environment-friendly technical route for industrial production of chiral alcohol products, and has very high industrial application potential.

The invention belongs to the technical field of gene engineering, and provides rice purple acid phosphatase OsPAP16 and application thereof, and the application is that a rice purple acid phosphatase OsPAP16 mutant is applied to rice cultivation. Two OsPAP16 mutants are prepared by adopting a gene editing technology, then the mutants are subjected to overexpression induction, and the result shows that the purple acid phosphatase OsPAP16 is positioned on a cell membrane, the OsPAP16 is subjected to phosphorus deficiency induced expression on leaves and roots, the phosphorus deficiency induced expression at the roots is quicker and higher, the overexpression of the OsPAP16 can improve the activity of the acid phosphatase on the root surface, and the yield of the acid phosphatase on the root surface is increased. Phosphorus deficiency stress of the rice can be relieved by degrading organic phosphorus in the environment, the phosphorus content of the rice is increased, and then growth and development of the rice are promoted.

The invention discloses application of a TpGST gene in regulating and controlling ear rot and stem rot resistance of maize fusarium graminearum, and belongs to the technical field of gene engineering. According to the application of the TpGST gene disclosed by the invention in regulating and controlling the ear rot and stem rot resistance of maize fusarium graminearum, the nucleotide sequence of the TpGST gene is shown as SEQ ID NO. 1. Fusarium graminearum is inoculated to ears and stems of an overexpressed TpGST transgenic line, so that the ear rot and stem rot resistance of the fusarium graminearum of the corn transgenic line is improved; fusarium graminearum is inoculated to ears and stems of the corn hybrid agricultural large 3138 overexpression TpGST transgenic line, and the ear rot and stem rot resistance of the fusarium graminearum of the corn transgenic hybrid is improved.