181 results about "Gene engineering" patented technology

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 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 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 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 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 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 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 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 belongs to the technical field of enzyme engineering and gene engineering, and particularly relates to a recombinant immobilized enzyme, an expression gene thereof and application of the recombinant immobilized enzyme in production of macrocyclodextrin. According to the preparation method, SpyTag and SpyCatcher peptides are fused to the N end of isoamylase and the N end of 4-alpha-glucanotransferase respectively, the 4-alpha-glucanotransferase modified by the SpyTag and the isoamylase modified by the SpyCatcher are obtained, and after spontaneous isopeptide bonds are formed between the SpyTag and the SpyCatcher, the self-assembled macrocyclodextrin production enzyme cluster is obtained. The obtained macrocyclodextrin production enzyme cluster forms an enzyme aggregate with a three-dimensional structure under the traditional glutaraldehyde-mediated catalytic action, namely the recombinant immobilized enzyme. The recombinant immobilized enzyme disclosed by the invention can be repeatedly utilized, is simple and convenient to operate, is easy to separate a converted product, and can be applied to industrial production of the macrocyclodextrin.

The application belongs to the technical field of genetic engineering, and particularly relates to application of a GadW mutant in human milk oligosaccharide production. The amino acid sequence of the GadW mutant is shown as SEQ ID NO. 3. By using a gene editing technology, a genetic engineering chassis strain carrying a GadW mutant coding gene is obtained, and then the above chassis strain is applied in a human milk oligosaccharide such as 3'-SL, 6'-SL, LNT II, LNnT and LNT, etc. production strain construction process, and the yield of human milk oligosaccharide is further improved. Among them, the yield of 3'-SL is increased by 8.5%, the yield of LNT II is increased by 26.7%, the yield of LNnT is increased by 50%, the yield of LNT is increased by 42.9%, and the yield of 6'-SL is increased by 35.3%.

The invention is applicable to the technical field of gene engineering, and provides application of an OsWRKY1 gene in improvement of saline-alkaline tolerance of rice, and the nucleotide sequence of the OsWRKY1 gene is as shown in SEQ ID No.1. The molecular mechanism of response of OsWRKY1 to saline-alkali stress is explored, a new theoretical basis is provided for improving saline-alkali tolerance of rice by using a molecular breeding means and cultivating a new variety of saline-alkali tolerant rice, and great significance is achieved for improving the yield and quality of the rice.

The present application relates to a kind of whole cell catalytic synthesis N-acetyl-trans-4-hydroxyproline method, belong to biological catalysis technical field.The present application is constructed by mutating acyltransferase and heterologous expression gene engineering bacteria, successfully realized using whole cell catalytic synthesis N-acetyl-trans-4-hydroxyproline, avoids the problem of enzyme instability, reduces the separation cost of enzyme and product.On this basis, the present application improves the yield of N-acetyl-trans-4-hydroxyproline by optimizing catalytic time, cell addition amount, the type and addition amount of additive, and the yield reaches 239mg / L.The present application is a kind of N-acetyl-trans-4-hydroxyproline preparation method with lower cost, lower equipment demand, more suitable for industrial production.

The invention discloses a PavSWEET9b gene for regulating and controlling the sugar content of sweet cherry fruits and application of the PavSWEET9b gene, and belongs to the technical field of gene engineering. The nucleotide sequence of the PavSWEET9b gene for regulating and controlling the sugar content of the sweet cherry fruits is shown as SEQ ID NO.1. The research finds that the PavSWEET9b protein is positioned on a cell membrane, and the soluble sugar content of the sweet cherry fruits is correspondingly reduced or increased after the gene is silenced and overexpressed, so that the PavSWEET9b gene can regulate and control the soluble sugar accumulation of the sweet cherry fruits; according to the method, the content of sugar in the sweet cherry fruits can be increased, so that the fruit quality is improved, the taste of the sweet cherry fruits is improved, the requirements of consumers are better met, and a new idea is provided for development of new sweet cherry varieties.

The invention provides application of an OsATG5 gene in improvement of drought resistance of rice, and belongs to the technical field of gene engineering and rice drought resistance. The nucleotide sequence of the OsATG5 gene is as shown in SEQ ID NO: 1, and the amino acid sequence of the encoding protein of the OsATG5 gene is as shown in SEQ ID NO: 2. Compared with a wild type (WT), the survival rate of the OsATG5 gene mutant plant osatg5 under PEG-induced drought stress is obviously lower than that of the WT; the survival rate of the plant OsATG5-OE overexpressed by the OsATG5 gene under drought stress is obviously higher than that of WT. Compared with WT, the content of proline and soluble sugar in the plant OsATG5-OE overexpressed by the gene OsATG5 after drought stress induced by PEG is increased, and the accumulation amount of hydrogen peroxide and malondialdehyde is reduced; after PEG induces drought stress, the proline content and the soluble sugar content of osatg5 mutant leaves are reduced, and the hydrogen peroxide content and the malondialdehyde content are increased. In conclusion, the OsATG5 gene is a potential candidate new target for regulating and controlling the drought resistance mechanism of the rice.

The present application belongs to the technical field of bioengineering, and particularly relates to a Claudin18.2 gene engineering antibody based on DNA immunization and a preparation method and application thereof. The Claudin18.2 gene engineering antibody comprises a heavy chain variable region and a light chain variable region; the heavy chain variable region comprises any one of the amino acid sequences shown in SEQ ID NO. 15-16; and the light chain variable region comprises the amino acid sequences shown in SEQ ID NO. 19-20. The Claudin18.2 gene engineering antibody is obtained by immunizing Balb / c mice with full-length plasmid DNA and full-length plasmid DNA+extracellular segment polypeptide as immunogens respectively, obtaining plasma cells capable of secreting specific antibodies against Claudin18.2, and then preparing specific mouse-derived gene engineering antibodies through single-cell optical guidance technology and genetic engineering technology. The gene engineering antibodies have high titer and strong specificity, and have specific recognition ability for natural protein Claudin18.2.

The invention discloses a populus euphratica PeXTH7 gene and application of the populus euphratica PeXTH7 gene in improving drought resistance of plants, and belongs to the technical field of gene engineering. The CDS sequence of the populus euphratica PeXTH7 gene is disclosed for the first time and is as shown in SEQ ID NO.1, and the coded amino acid sequence is as shown in SEQ ID NO.2; the effect of the populus euphratica PeXTH7 gene in improving the drought resistance of the plant is disclosed for the first time, the root length and lateral root density of the plant are remarkably improved through overexpression of the populus euphratica PeXTH7 gene, and the drought resistance of the plant is improved by adjusting the root structure of the plant and promoting root development; the populus euphratica PeXTH7 gene provided by the invention can be used as a candidate gene for breeding the drought-resistant new variety of the plant, and is of great significance for cultivating the drought-resistant new variety of the plant by adopting a genetic engineering method, accelerating the breeding process and reducing the breeding workload.

The invention relates to the technical field of gene engineering, and particularly discloses a chitinase truncated body, a recombinant expression vector thereof, and a construction method and application of engineering bacteria. The chitinase truncated body is Cq181NC, the amino acid sequence of the Cq181NC is as shown in SEQ ID NO: 1, and the nucleotide sequence for coding the chitinase is as shown in SEQ ID NO: 2. According to determination, the chitinase truncation provided by the invention has the advantages that the specific enzyme activity is obviously improved by 1.8 times compared with that of a wild type, the chitinase truncation is high in environmental change tolerance, and the thermal stability of the chitinase truncation is still 1.43 times that of the wild type (incubation is performed for 8 hours at 35 DEG C). More importantly, the spectrum of the enzymolysis product is changed: the wild type is mainly disaccharide and shows a typical excision enzyme mode; and the truncated Cq181NC generates a plurality of oligosaccharides including monosaccharide, disaccharide, trisaccharide and hexasaccharide, shows double catalytic characteristics of endonuclease and exonuclease, and has a better application prospect.

The invention relates to a primer group for detecting Rafivirus, a kit and application, and relates to the technical field of gene engineering. The primer group comprises a forward primer and a reverse primer, the forward primer is selected from SEQ ID NO: 2, 3 and 4, and the reverse primer is selected from SEQ ID NO: 5, 6 and 7. On the basis of the primer group, Rafivirus can be specifically recognized, so that the primer group is combined with an RAA amplification system and a detection system, and detection is rapidly completed in a short time under the condition of constant temperature.

The invention belongs to the technical field of gene engineering, and particularly relates to a CHO cell transient transfection method and application thereof.The CHO cell transient transfection method comprises the following steps that subcultured CHO cells are resuspended through a transfection culture medium, the cell density in CHO cell suspension is 0.8 * 10 < 7 >-1.5 * 10 < 7 > cells / mL, a PEI transfection reagent containing 40 kDa linear PEI is used, plasmid DNA and the PEI transfection reagent with the mass ratio being 1: (1-6) are added into the CHO cell suspension for transfection, and transfection is conducted after the PEI transfection reagent is added into the CHO cell suspension; the addition amount of plasmid DNA in each milliliter of CHO cell suspension is 3.0-7.0 [mu] g, transfecting for 20-24 h, then adding a cell suspension culture medium, transfecting for 48-72 h, then repeatedly transfecting, and culturing to obtain the recombinant CHO cell. According to the method, the transfection efficiency can be improved, more cells for expressing the recombinant protein can be obtained, and the transient expression of the target protein can be remarkably improved.