259 results about "Gene engineering" patented technology

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 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 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 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 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 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 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 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.

The invention discloses a keto isomerase mutant as well as a preparation method and application thereof, and relates to the technical field of gene engineering. The keto isomerase mutant is obtained by performing the following mutations on an amino acid sequence shown as SEQ ID NO.2: the 34th amino acid is mutated from N to R; meanwhile, the 188th amino acid is mutated from I to K. The half-life period of the keto isomerase mutant obtained by mutating the amino acid sequence shown in SEQ ID NO.2 is prolonged, and the enzyme activity is improved, so that the yield of D-chiro-inositol is further improved.

The invention relates to the field of gene engineering and molecular breeding, and discloses application of a ZmTCP5 gene in regulation and control of heat resistance of corn. According to the invention, three types of knockout mutants of the gene are obtained by using a CRISPR / Cas9 technology, and three overexpression materials are created. At normal temperature (25-28 DEG C), the material has no obvious phenotypic difference with a wild type; however, under high temperature stress (45 DEG C), the ZmTCP5 knockout mutant has enhanced heat resistance, reduced reactive oxygen species (ROS) accumulation, and significantly increased ROS scavenging enzyme activity, chlorophyll content and photochemical efficiency; on the contrary, the ZmTCP5 overexpression strain is more sensitive to heat stress, which indicates that the ZmTCP5 negatively regulates the heat resistance of the corn. The research provides a theoretical basis for analyzing a heat-resistant molecular mechanism of the corn, and also provides an important gene resource for creating a new germplasm of the heat-resistant corn.

The invention belongs to the technical field of gene engineering, and particularly relates to fusion vitamin D hydroxylase and application thereof in preparation of 25-hydroxy vitamin D2 and 25-hydroxy vitamin D3. The fusion vitamin D hydroxylase comprises a Vdh-K1 heme structural domain of which the amino acid sequence is shown as SEQ ID NO.1, a CYP116B65 redox structural domain of which the amino acid sequence is shown as SEQ ID NO.3, and a linker for connecting the Vdh-K1 heme structural domain and the CYP116B65 redox structural domain. The fusion vitamin D hydroxylase is a self-sufficiency enzyme, and can realize stable catalytic conversion of vitamin D2 and vitamin D3 on the premise of not depending on an Fdx and Fdr assisted electron transfer system.

The invention discloses an application of an SCAB82331 gene or a protein coded by the SCAB82331 gene in regulation and control of synthesis of thaxtominA, and belongs to the technical field of gene engineering. According to the application disclosed by the invention, the SCAB82331 gene in the streptomyces scabies is deleted through a genetic engineering way, so that the high-yield strain of the thaxtominA is obtained, and the yield of the thaxtominA in the streptomyces scabies is increased. When the SCAB82331 gene is deleted in the streptomyces scabies, the yield of the thaxtominA is obviously increased by 66.2%; after the SCAB82331 gene is supplemented in the delta SCAB82331 mutant strain, the yield of the thaxtominA is recovered, and the yield of the thaxtominA is recovered; the deletion of the SCAB82331 gene can be used for directionally improving the biosynthesis yield of the thaxtominA in streptomyces scabies.

The invention discloses a rapid detection method for traumatic brain injury by fusing genetic engineering and quantum dots, and relates to the technical field of traumatic brain injury detection. The method comprises the following steps: acquiring a suspected patient biological sample, extracting biomolecule components, and comparing the biomolecule components with a healthy population biomolecule database to obtain related differential expression molecules. A specifically recognizable recombinant antibody is constructed by genetic engineering, and is connected with a quantum dot through a coupling reaction to prepare a detection probe. The probe and a biological sample are mixed and reacted, and the intensity of a fluorescence signal generated by combination is collected by a fluorescence detection device. A standardized signal value is obtained through signal processing algorithm noise reduction and feature extraction, and a preliminary detection result is determined by combining a bioinformatics database evaluation sample. And finally, according to the differential expression molecule, the standardized signal value and the preliminary result, analyzing a data change trend in a preset time window by using a multivariable statistical model, and generating a diagnosis report. According to the method, various technologies are fused, and a new path is provided for traumatic brain injury detection.

The invention belongs to the technical field of gene engineering and protein engineering, and particularly relates to thermal-stable D-psicose 3-epimerase as well as a preparation method and application thereof. According to the invention, a series of D-psicose 3-epimerase DAE mutants are constructed, and six mutants which not only can retain good catalytic activity, but also have high thermal stability are obtained through screening. The catalytic temperature of the three mutants A19D / A223P / V241I is increased by 25 DEG C compared with that of a wild type, while the three mutants A19D / A223P / V241I still show the optimal thermal stability, the catalytic activity of the three mutants is improved by 17.0%, and the three mutants show good industrial suitability. The D-psicose 3-epimerase DAE mutant with improved thermal stability provided by the invention is beneficial to improving the production efficiency of the DAE in industrial application of D-psicose, and provides powerful industrial enzyme guarantee for large-scale and low-cost preparation of D-psicose.

The invention provides application of GmEDS5a and / or GmEDS5b genes in regulation and control of soybean root nodule size and soybean nitrogenase activity or in soybean breeding, and belongs to the technical field of gene engineering. The GmEDS5a and / or GmEDS5b genes are / is knocked out, the proportion of root nodules with the particle size larger than 2 mm is increased, the soybean nitrogenase activity is remarkably improved, symbiotic nitrogen fixation is promoted, and great significance is achieved for increasing the soybean yield.

The invention relates to the technical field of gene engineering, in particular to a grape transcription factor VvNAC53, application of the grape transcription factor VvNAC53 and a method for regulating and controlling the content of terpenoids in grapes. The nucleotide sequence for coding the grape transcription factor VvNAC53 is as shown in SEQ ID NO. 1; the content of terpenoids in grapes can be regulated and controlled by adopting VvNAC53; the regulation and control method comprises the following steps: transferring a grape transcription factor VvNAC53 into grapes and performing overexpression to reduce the content of terpenoids in grape fruits so as to directionally cultivate grape germplasm with different aroma types.

The invention belongs to the technical field of gene engineering, and relates to application of a receptor-like kinase gene MdCIK2 in regulation and control of apple disease resistance. The invention provides an application of a receptor-like kinase gene MdCIK2 in regulation and control of apple disease resistance, which comprises the following steps: a) silencing the receptor-like kinase gene MdCIK2 to improve the resistance of apples to rot diseases; or b) silencing a specific gene segment of the receptor-like kinase gene MdCIK2, and improving the resistance of apples to rot disease. The receptor-like kinase gene MdCIK2 for negative regulation of apple disease resistance provided by the invention can provide important gene resources for apple disease resistance molecular breeding and targeted gene editing.

The invention provides application of a rice NAC028 gene in regulation and control of rice aluminum resistance, and belongs to the technical field of gene engineering. The invention provides an application of an NAC028 protein and / or a gene for coding the NAC028 protein in regulating and controlling the tolerance of a plant to aluminum. A gene function-deleted mutant nac028 plant and an overexpression plant are created and are subjected to aluminum stress treatment together with wild Nipponbare, and the result shows that the gene-deleted mutant is super sensitive to aluminum, and the tolerance of the overexpression plant to aluminum is enhanced. The NAC028 protein or the coding gene of the NAC028 protein can be used for perfecting an upstream and downstream regulation and control network of rice responding to aluminum stress, and also provides an important gene resource and a new path for breeding of aluminum-resistant plants.

The invention provides an alginate lyase mutant Amut8C as well as a coding gene and application thereof, belongs to the technical field of gene engineering and enzyme engineering, and can solve the problem that existing alginate lyase cannot meet the requirements of severe reaction conditions such as high temperature and acidity in industrial application. The amino acid sequence of the alginate lyase mutant Amut8C disclosed by the invention is as shown in SEQ ID NO. 1. According to the space structure of the alginate lyase Algt, disulfide bonds in molecules are analyzed, the alginate lyase is mutated, and the obtained alginate lyase mutant Amut8C has heat resistance, acid and alkali resistance and digestive enzyme tolerance, and compared with the alginate lyase Algt, the alginate lyase mutant Amut8C has the advantages that the content of the alginate lyase mutant Amut8C is increased, and the content of the alginate lyase mutant Amut8C is increased. The alginate oligosaccharide with the polymerization degree of 1-4 prepared from the alginate lyase mutant Amut8C under the same enzymolysis condition is higher in yield. The method can be applied to the fields of feeds, foods, detergents or biofuels.

The invention discloses a ZaSCL3 gene for regulating and controlling plant growth and development as well as an encoding protein and application thereof, and belongs to the technical field of gene engineering. According to the invention, an over-expression vector of the ZaSCL3 gene is constructed and expressed in a wild tomato, so that stamens of a ZaSCL3 over-expression plant are aborted, ovary is spontaneously expanded without pollination, and seedless fruits are generated, which indicates that the over-expression ZaSCL3 gene can realize the effect of inducing male sterility and apomixis of the plant; a key gene resource is provided for molecular breeding and yield character improvement, and the important application value is achieved.

The invention discloses an acid protease mutant PepA4 with improved heat resistance and application thereof, and relates to the technical field of gene engineering and enzyme engineering. The amino acid sequence of the acid protease mutant PepA4 is obtained by carrying out D67A, E103D, E304K and S368L point mutation on an amino acid sequence as shown in SEQ ID NO: 1, and the amino acid sequence of the acid protease mutant PepA4 is as shown in SEQ ID NO: 2. The high-temperature-resistant acid protease disclosed by the invention can still maintain the enzyme activity of 80% or above at 75 DEG C for 5 minutes. The acid protease provided by the invention has better acid resistance, and can be widely applied to food processing, leather processing and feed processing industries.

The invention belongs to the technical field of gene engineering and cell transformation, and discloses a gene engineering construction method and application of anti-apoptosis human mesenchymal stem cells. Through HDAC1 K412 site specific lactic acid mutation, breakthrough improvements are realized: for example, CRISPR-Cas9-mediated accurate gene editing is utilized, only a single amino acid site is changed, that is, K412R mutation simulates persistent lactic acid, and whole genome apparent modification disturbance can be avoided; the modification does not depend on an exogenous metabolic substrate, and the negative feedback effect of traditional metabolic intervention is overcome; the mutant continuously inhibits a P53 apoptosis pathway, so that the oxidative stress mediated apoptosis resistance of the MSC is remarkably improved. According to the design concept of one target spot and one regulation, the core contradiction that long-term effectiveness and specificity cannot be considered in the prior art is fundamentally solved, and a brand new normal form is provided for stem cell treatment.

The invention discloses an alpha-1, 4 glucosidic bond hydrolytic ability enhanced oligomerized 1, 6-glucosidase mutant and application thereof, and belongs to the technical field of gene engineering and enzyme engineering. The oligomerized 1, 6-glucosidase mutant V219A / P227A disclosed by the invention also has the alpha-1, 4 glucosidic bond hydrolytic activity which is not possessed by a wild type enzyme on the basis of retaining the original alpha-1, 6 glucosidic bond hydrolytic activity, and the alpha-1, 4 glucosidic bond hydrolytic activity is improved by 267% compared with that of a single mutant V219A. The mutant V219A / P227A is applied to an enzymatic production process of glucose, maltodextrin is taken as a substrate, glucamylase and the mutant V219A / P227A are added for enzymatic conversion, and the final glucose yield reaches 98.0%; the mutant V219A / P227A is applied to regeneration of glucose from crystalline glucose mother liquor, and the final glucose yield reaches 95.9%, which is 9.35% and 5.15% higher than those of a wild type V219A and a single mutant V219A respectively. Therefore, the mutant is helpful for improving the substrate conversion rate and the product purity in the glucose production process, can be used for regenerating glucose from crystalline glucose mother liquor, and has higher industrial application value.