2939 results about "Expression vector" patented technology

The invention belongs to the technical field of gene engineering and enzyme engineering, and particularly relates to an ornithine decarboxylase mutant as well as a coding gene, an expression vector, a recombinant strain and application thereof. The ornithine decarboxylase mutant is obtained by mutating wild ornithine decarboxylase ODC10 of which the amino acid sequence is shown as SEQ ID NO.1, the catalytic activity of the obtained mutant is improved, and the biosynthesis efficiency of 1, 4-butanediamine is favorably improved. The recombinant strain for expressing the ornithine decarboxylase mutant constructed by the invention can efficiently synthesize 1, 4-butanediamine, and the molar conversion rate of ornithine is remarkably improved, so that the production benefit is improved, and the production cost is reduced.

The present invention relates to the fields of genetic engineering and plant genetic breeding, and specifically to the application of wheat TaPND gene. In the present invention, by taking gene TaPND as a target, designing a sgRNA sequence based on CRISPR / Cas9, connecting a DNA fragment containing the encoding sgRNA sequence to a vector carrying CRISPR / Cas, a plant binary expression vector capable of knocking out the TaPND gene in wheat is constructed; by genetically transforming wheat, offspring with TaPND gene knockout are obtained, and wheat mutants with early flowering time are obtained. Experiments have confirmed that transgenic wheat with TaPND gene knockout achieves the advancement of wheat flowering time. The technical scheme of the present invention provides a practical method for realizing rapid wheat breeding using genetic engineering technology, which has important breeding application value and broad market application prospects.

Provided herein are engineered nucleic acids (e.g., expression vectors, including viral vectors, such as lentiviral vectors, adenoviral vectors, AAV vectors, herpes viral vectors, and retroviral vectors) that encode OCT4; KLF4; SOX2; or any combination thereof that are useful, for example, in inducing cellular reprogramming, tissue repair, tissue regeneration, organ regeneration, reversing aging, or any combination thereof. Also provided herein are recombinant viruses (e.g., lentiviruses, alphaviruses, vaccinia viruses, adenoviruses, herpes viruses, retroviruses, or AAVs) comprising the engineered nucleic acids (e.g., engineered nucleic acids), engineered cells, compositions comprising the engineered nucleic acids, the recombinant viruses, engineered cells, engineered proteins, chemical agents that are capable of activating expression of OCT4; KLF4; SOX2; or any combination thereof, an engineered protein selected from the group consisting of OCT4; KLF4; SOX2; or any combination thereof, an antibody capable of activating expression of OCT4; KLF4; SOX2; or any combination thereof, and methods of treating a (e.g., ocular disease), preventing a disease (e.g., ocular disease), regulating (e.g., inducing or inducing and then stopping) cellular reprogramming, regulating tissue repair, regulating tissue regeneration, or any combination thereof).

The invention discloses a mammalian cell expression vector, an expression plasmid and an expression system. The expression vector sequentially comprises the following elements: (a) a promoter; (b) a Kozak sequence; (c) a polyadenine sequence, the polyadenine sequence sequentially comprises a bGH PolyA sequence and a 5 'UTR PolyA sequence, the nucleotide sequence of the bGH PolyA sequence is as shown in SEQ ID NO.5, and the nucleotide sequence of the 5' UTR PolyA sequence is as shown in SEQ ID NO.6; (d) copying the start site; and (e) screening marker genes. According to the present invention, the expression vector is optimized by the expression element, such that the exogenous gene can be conveniently, efficiently and flexibly inserted and edited by the expression vector so as to achieve the stable extracellular or intracellular expression of the exogenous protein, and the expression efficiency of the exogenous protein can be improved by carrying out the expression plasmid construction based on the expression vector; and more efficient expression and purification of the foreign protein are realized.

The invention discloses an FtACA13 gene derived from tartary buckwheat as well as an expression vector and application of the FtACA13 gene. According to the invention, the FtACA13 gene is separated from tartary buckwheat; the polynucleotide sequence of the FACA13 gene is as shown in SEQ ID NO.1, and the amino acid sequence of the encoded protein of the FACA13 gene is as shown in SEQ ID NO.2. The FtACA13 gene is genetically transformed into buckwheat to obtain transgenic buckwheat hairy roots with the overexpressed FtACA13 gene, compared with hairy roots of a control group, the growth vigor and fresh weight of the fagopyrum tataricum hairy roots with the overexpressed FtACA13 gene under salt stress are increased, Na < + > accumulation in a plant body is remarkably low, it is proved that the FtACA13 gene can regulate and control the salt stress resistance of the plant, and the salt stress resistance of the fagopyrum tataricum hairy roots is improved. The invention has application prospects in the aspects of cultivating or breeding salt stress resistant plant varieties and the like.

The present invention relates to the field of biotechnology and medicine, in particular to a monoclonal antibody or antigen-binding fragment thereof that specifically binds to TNF-like ligand 1A (TL1A). The invention further relates to nucleic acids encoding said antibody, expression vectors, host cells and methods for producing same, methods for producing the antibodies according to the invention, pharmaceutical compositions comprising the antibody or antigen-binding fragment thereof according to the invention, as well as to pharmaceutical compositions comprising the antibody or antigen-binding fragment thereof according to the invention and other therapeutically active compounds; to methods for treating TL1A-mediated diseases or disorders, to use of the antibody or antigen-binding fragment thereof or pharmaceutical composition thereof for treating TL1A-mediated diseases or disorders, and to use of the antibody or antigen-binding fragment thereof according to the invention and other therapeutically active compounds for treating TL1A-mediated diseases or disorders.

The invention discloses a Chinese orchid flowering regulation gene CsFTL3 as well as an encoding protein and application thereof, and belongs to the field of plant molecular biology. The gene CsFTL3 for regulating and controlling flowering of Chinese cymbidium is separated from flower bud cDNA (complementary deoxyribonucleic acid) of a Chinese cymbidium variety 'Xiaoxiang', and 174 amino acids are encoded by the gene CsFTL3. Through construction of an expression vector of the CsFTL3 gene, overexpression in wild-type arabidopsis thaliana and heterologous overexpression of the CsFTL3 gene in the arabidopsis thaliana, it is found that bolting and flowering periods of the arabidopsis thaliana are significantly delayed, flower stems are significantly thickened and increased, the number of rosette leaves is significantly increased, and the growth period is prolonged, which indicates that the gene has the function of delaying the flowering time of the arabidopsis thaliana. The Chinese orchid flowering regulation and control gene can be used for molecular mechanism research of orchid flowering time, delay Chinese orchid flowering time, promote plant vegetative growth, prolong whole Chinese orchid viewing time and promote Chinese orchid genetic improvement and variety breeding.

The invention discloses an application of a rape BnC05ERF378 gene in improving the waterlogging resistance of a plant. The nucleotide sequence of the rape BnC05ERF378 gene is as shown in SEQ ID NO. 1. According to the invention, a brand new stain-resistant gene BnC05ERF378 is cloned from rape for the first time, and is transferred into arabidopsis thaliana to construct a pure line plant and complete stain resistance identification. Experimental results show that the growth state of the transgenic BnC05ERF378 gene arabidopsis thaliana is remarkably superior to that of a wild type after waterlogging treatment, the transgenic BnC05ERF378 gene arabidopsis thaliana can normally bolt, the survival rate is increased from 12.5% to 37.5%, it is fully proved that the gene can enhance the survivability of the plant in a waterlogging environment by regulating the response mechanism of the plant to waterlogging stress, and the survival rate of the plant is increased from 12.5% to 37.5%. And a key guarantee is provided for stable growth of plants under high-humidity, waterflooding and other adverse conditions. According to the discovery and application of the waterlogging-resistant gene BnC05ERF378 disclosed by the invention, a clear functional gene target is provided for improvement of waterlogging resistance of crops. By constructing an expression vector containing the gene and combining mature technologies such as agrobacterium transformation, the gene can be stably introduced into a target plant, and a pure transgenic plant is obtained.

The invention discloses a method for improving citrus yellow vein disease resistance by using a ClPsaN gene. The method comprises the following steps: (1) cloning a VIGS fragment of the citrus ClPsaN gene; (2) constructing two VIGS expression vectors of TRV and CLBV; and (3) transforming the citrus by the VIGS expression vector to obtain a citrus ClPsaN gene silenced VIGS plant. According to the invention, the VIGS vector of the Citrus ClPsaN gene is constructed, and then the Citrus is transformed, so that the obtained Citrus plant can show obvious resistance to the Citrus yellow vein clearing virus, the titer of the Citrus yellow vein clearing virus can be reduced by 22-34%, the accumulation amount of the virus protein is reduced by 35-50%, and the attack degree of the Citrus yellow vein clearing virus is obviously reduced; the TRV silencing of the citrus ClPsaN gene does not influence the phenotype of a citrus plant, and the citrus ClPsaN gene can be used as a candidate gene for citrus yellow vein disease resistance breeding together with a plurality of citrus yellow vein disease-resistant and susceptible genes by utilizing the cooperation of VIGS silencing, RNA interference, gene editing and other technologies.

The invention relates to the technical field of biology, and particularly discloses a recombinant collagen III and application thereof in preparation of gel. The recombinant collagen III is designed by optimizing functional area sequences of human I-type and III-type collagen, the amino acid sequence is shown as SEQ ID No.2, and the recombinant collagen III has the characteristics of high stability, good hydrophilicity and low immunogenicity. The preparation method comprises the steps of expression vector construction, escherichia coli induced expression, affinity chromatography purification and renaturation. The recombinant collagen III can be prepared into a gel dressing and comprises sodium alginate, methylparaben and other components. Experiments show that the gel can effectively promote cell proliferation and has no cytotoxicity; in a mouse skin injury model, the collagen can relieve inflammation, accelerate wound healing and inhibit scar formation, and the effect of the collagen is superior to that of natural human III-type collagen. The invention provides a safe and efficient novel material for wound repair, and is suitable for medical dressings and tissue engineering.

The invention discloses a high-efficiency lysine decarboxylase mutant and an application of the high-efficiency lysine decarboxylase mutant in synthesis of pentamethylene diamine. A series of mutants including a single mutant (such as N218G, T222V, C244P and the like), a double mutant and a three mutant are obtained by carrying out multi-site mutation on wild type lysine decarboxylase (SEQ ID NO.2) from Klebsiella grimontii. The catalytic efficiency of the mutant is remarkably improved, and the activity of the mutant is improved by 5.0-10.2 times compared with that of a wild type enzyme. According to the method disclosed by the invention, the substrate conversion rate of 99% or above can be realized within 2 hours through an optimized process of constructing an expression vector and a genetically engineered bacterium, inducing expression of lysine decarboxylase and synthesizing 1, 5-pentanediamine by utilizing a whole-cell catalysis technology. Compared with a traditional chemical method and a natural enzyme method, the method disclosed by the invention has the advantages of simplified process, high catalytic efficiency, low production cost and the like, and is suitable for producing bio-based 1, 5-pentamethylene diamine and nylon 56 salt.

The invention belongs to the technical field of gene engineering, and relates to a cyperus esculentus CeMYB154 gene capable of improving salt tolerance and active oxygen scavenging capacity and application of the cyperus esculentus CeMYB154 gene. The Cyperus esculentus CeMYB154 gene belongs to an MYB transcription factor family, the Cyperus esculentus CeMYB154 gene is screened and transformed into Arabidopsis thaliana through transcriptome sequencing, bioinformatics analysis, gene cloning, overexpression vector construction and recombinant strain overexpression, and analysis finds that under salt stress, compared with a wild type, the root length of a plant with the overexpressed CeMYB154 gene is obviously increased, the growth vigor is better, and the growth rate of the plant with the overexpressed CeMYB154 gene is higher. According to the present invention, the content of MDA and H2O2 is significantly reduced, and the activity of CAT, POD and SOD antioxidant enzymes is significantly enhanced, such that the overexpressed CeMYB154 gene can enhance the salt tolerance and the active oxygen scavenging ability of the plant, and the overexpressed CeMYB154 gene is proved to be the salt tolerance positive regulation factor; the CeMYB154 gene can be used in the field of salt-tolerant stress-resistant breeding of cyperus esculentus, and provides an important gene resource for salt-tolerant molecular breeding.

The invention discloses an aspergillus niger recombinant bacterium AnCT-XynB-K-agdA and application thereof, and belongs to the technical field of microbiology and bioengineering. According to the invention, Aspergillus niger AnCat is taken as an expression host, firstly, an acid protease regulatory factor prtT is knocked out to obtain a defective strain, and then different chaperonins are subjected to fusion expression by optimizing integration sites of a laccase lcc9 expression cassette, so that the high-expression laccase chaperonin is obtained. According to the method, an expression vector pC3-5 'agdA-PcitA-XynB-L-lcc9-hph-3' agdA is constructed, a recombinant bacterium AnCT-XynB-K-agdA is further obtained, the enzyme activity of the extracellular laccase subjected to shake flask fermentation reaches 1821.2 U / L, and finally, the recombinant efficient expression of the laccase Lcc9 in aspergillus niger is realized.

The invention belongs to the technical field of living cell imaging, and discloses a living cell nucleic acid imaging system and method based on CRISPR (clustered regularly interspaced short palindromic repeats) and a nano antibody and application, the system comprises: a modified dCas protein expression vector containing n antigen polypeptide tags; the modified sgRNA and / or crRNA expression vector comprises a specific guide sequence; the fluorescent nano-antibody expression vector contains m fluorescent nano-antibody coding sequences which are connected in an operable manner, and fluorescent protein serving as an internal tag is inserted between nano-antibody splitting sites, can be combined with a polypeptide tag carried by dCas protein and can stably emit light; and the fluorescent nano antibody in a free state can be self-degraded, so that the fluorescent nano antibody does not emit fluorescence. According to the invention, the single-copy nucleic acid sequence, especially the imaging marker of the single-copy nucleic acid sequence in a living cell, can be visualized with single-molecule resolution and high signal-to-noise ratio.

The invention discloses a pinus massoniana PmYTH1 gene as well as an encoding protein and application thereof, and relates to the technical field of plant genetic engineering. The nucleotide sequence of the pinus massoniana PmYTH1 gene disclosed by the invention is as shown in SEQ ID NO. 1. The method comprises the following steps: constructing an expression vector of a pinus massoniana PmYTH1 gene, and transforming the expression vector into an arabidopsis thaliana mutant; and cultivating and screening to obtain a transgenic arabidopsis thaliana plant of which the ABA tolerance is obviously increased. The result shows that the tolerance of the arabidopsis thaliana ect2 / 3 / 4 mutant to ABA is improved by overexpressing the PmYTH1 in the arabidopsis thaliana ect2 / 3 / 4 mutant; the expression quantity of ABA signal channel genes is obviously increased. The mRNA of the ABA signal channel gene in the 35S:: PmYTH1 / ect2 / 3 / 4 strain is more stable. The invention provides a molecular means and basis for cultivating a new pinus massoniana variety with high ABA and adversity stress tolerance.

The invention belongs to the technical field of plant genetic engineering molecular breeding, and particularly relates to application of wheat TabHLH93 protein or a coding gene thereof in regulation and control of plant grain development. According to the invention, a binary expression vector capable of realizing ectopic expression of the TabHLH93 gene is constructed and is genetically transformed into common wheat to obtain a progeny with high expression of the TabHLH93 gene, so that overexpression of the TabHLH93 gene is realized, and it is verified that the TabHLH93 has a function of regulating and controlling the size of wheat grains and also has an important regulation and control effect on agronomic characters such as wheat plant height, tiller number, spike length and thousand seed weight. A candidate gene with an important application prospect is provided for wheat high-yield genetic improvement, and the gene has important significance for guaranteeing grain safety.

The invention discloses a tomato LACS2 gene promoter and application thereof, and belongs to the technical field of biology. On one hand, the invention provides the tomato LACS2 gene promoter, and on the other hand, the invention provides application of the tomato LACS2 gene promoter in promoting target gene expression in plants. According to the invention, a tomato LACS2 gene promoter fragment (788bp) is constructed to drive an EGFP-GUS (Enhanced Green Fluorescent Protein-Glucuronidase) gene fusion expression vector. GUS histochemical staining and laser confocal microscopy observation prove that the promoter fragment can guide normal expression of the reporter gene in the tomato epidermis. Therefore, the LACS2 promoter can be used for researching development, quality improvement and postharvest preservation of tomato peels and specific expression of exogenous genes in the tomato peels.

The invention relates to the field of animal breeding, and discloses a pig beta defensin compound preparation which comprises pig beta defensin PBD1, PBD2 and PBD114, and the weight ratio of the PBD1 to the PBD2 to the PBD114 is 1: 1: 1. The porcine beta defensin PBD1, the porcine beta defensin PBD2 and the porcine beta defensin PBD114 are recombinant proteins, and are respectively expressed in escherichia coli BL21 (DE3) PLysS competent cells by a pET-32a expression vector. Through the synergistic effect of the defensins PBD1, PBD2 and PBD114, the minimum inhibitory concentration (MIC) of the EPEC is reduced to 18.75 mu g / mL from 75 mu g / mL of single defensins, and is reduced by 75%; within 24 hours, the bacteriostasis rate of the compound preparation reaches 90%, and the inhibition effect on EPEC is remarkably improved.

The invention belongs to the technical field of molecular biology, and particularly relates to a CeNF-YA3 gene, an expression vector and application of the CeNF-YA3 gene in vegetable fat regulation and control. The nucleotide sequence of the CeNF-YA3 gene is as shown in SEQ ID NO.1. After the CeNF-YA3 gene is over-expressed, the grease content of arabidopsis seeds and leaves can be remarkably increased; in addition, the invention also provides a series of vectors containing the CeNF-YA3 gene for subcellular localization, yeast hybridization and plant overexpression for the first time, and proves that the CeNF-YA3 protein has a transcriptional activation function, is localized in a cell nucleus and accords with the basic characteristics of transcription factors for the first time. Therefore, the technical scheme of the invention provides scientific guidance for the application of the CeNF-YA3 gene and the protein thereof.

The invention relates to cloning of a stress-resistant gene LpbHLH144 of Lilium pumilum DC (Lilium pumilum DC.) and application of the stress-resistant gene LpbHLH144 of Lilium pumilum DC. Comprising the following steps: (1) cloning a target gene; (2) constructing an expression vector; and (3) detecting physiological indexes of the transgenic plant. According to the experiment, lilium tenuifolium is taken as a research object, and the LpbHLH144 gene is cloned by applying an RT-PCR technology; transforming tobacco by constructing an overexpression vector; the functions of overexpression of the LpbHLH144 for improving the saline-alkaline and drought stress of the tobacco are identified through transgenic tobacco, the functions of the lilium tenuifolium bHLH gene when the lilium tenuifolium bHLH gene is subjected to adversity stress are disclosed, a foundation is laid for further application of the lilium tenuifolium in lily breeding, and a theoretical basis is provided for the functions and effects of bHLH transcription factors.

In some aspects, the disclosure relates to compositions and methods useful for inhibiting SOD1 expression in cells (e.g., cells of a subject). In some embodiments, the disclosure describes isolated nucleic acids engineered to express an inhibitory nucleic acid targeting endogenous SOD1 and an mRNA encoding a hardened SOD1 protein. In some embodiments, compositions and methods described by the disclosure are useful for treating Amyotrophic Lateral Sclerosis (ALS) in a subject.

The invention provides a subunit fusion protein RBD-Fc on the surface of a porcine D-type coronavirus and a preparation method and application thereof, the preparation method comprises the following steps: 1) cloning a gene sequence as shown in SEQ ID NO.5 into a eukaryotic expression vector to obtain a recombinant plasmid containing a fusion protein RBD-Fc coding gene; (2) transfecting a recombinant plasmid containing the fusion protein RBD-Fc coding gene into an expression cell; 3) culturing, screening and domesticating the expression cells in the step 2) to obtain a highly expressed cell strain; and 4) fermenting and culturing the cell strain in the step 3), and purifying to obtain the subunit fusion protein RBD-Fc. According to the present invention, the RBD-Fc can be subjected to mass soluble expression, the protein is stable, the problems that the porcine D-coronavirus surface S protein cannot be subjected to large-scale expression and the like in the prior art are solved, the preparation method is simple, and the cost is low.

The invention provides application of a porcine parainfluenza virus type 5 as an expression vector, and belongs to the technical field of biological vaccines. According to the invention, a PIV5 strain separated from diarrhea pigs in Jiangsu Province in China is utilized to construct PIV5 virus infectious clone. The eGFP gene is inserted into a PIV5 infectious cloned virus genome, the recombinant PIV5 virus capable of expressing the green fluorescent protein is successfully rescued, and when the eGFP gene is inserted into a P / M gene spacer region, the expression quantity is the highest, which indicates that an exogenous gene expression cassette can influence the expression level of an exogenous gene, and the P / M gene spacer region is the optimal site for inserting the exogenous gene, so that the recombinant PIV5 virus can be successfully rescued. And a solid foundation is laid for developing a vector vaccine by utilizing the swine PIV5. The porcine parainfluenza virus type 5 can be used as an expression vector to express foreign proteins, can also be used for preparing oral vaccines, and can relieve the stress reaction of pigs in the immune process.

The present disclosure relates to the seminal discovery of a generation and use of genetically engineered Vibrio sp. Provided is the use of the genetically engineered bacteria for the construction, maintenance, manipulation, and / or propagation of DNA constructs; protein expression; protein secretion; vectors and other metabolic tools; metabolic engineering; expression of cellular extracts for cell-free biology; shuttle vectors; cloning vectors; and for synthetic biology applications. The disclosure also relates to the use of the replication machinery of Vibrio sp. as a cloning or expression vector for replication of recombinant DNA constructs. The disclosure also relates to methods of use of the above.

The invention provides a recombined protein rSp14-3-3 of sarcocystis tenella, an antibody and an ELISA (Enzyme-Linked Immunosorbent Assay) diagnostic kit of the recombined protein rSp14-3-3 of the sarcocystis tenella. The research firstly provides the recombined protein rSp14-3-3 of the sarcocystis tenella. According to the present invention, the recombinant protein rSp14-3-3 is used as a gene fragment, the obtained gene fragment is connected to an expression vector to carry out induced expression and protein purification, the purified recombinant protein rSp14-3-3 is used to coat an ELISA plate, the ELISA detection method for sheep infected with sarcosporidium is established, and the specific monoclonal antibody for the recombinant protein rSp14-3-3 is further provided. All the products can be applied to diagnosis of infection of sarcocystis in samples, water samples, food, environmental samples and the like, and technical support is provided for subsequent immunological diagnosis of sheep sarcocystis by using the protein.

Provided herein are engineered nucleic acids (e.g., expression vectors, including viral vectors, such as lentiviral vectors, adenoviral vectors, AV vectors, herpes viral vectors, and retroviral vectors) that encode OCT4; KLF4; SOX2; or any combination thereof that are useful, for example, in inducing cellular reprogramming, tissue repair, tissue regeneration, organ regeneration, reversing aging, or any combination thereof. Also provided herein are recombinant viruses (e.g., lentiviruses, alphaviruses, vaccinia viruses, adenoviruses, herpes viruses, retroviruses, or AAVs) comprising the engineered nucleic acids (e.g., engineered nucleic acids), engineered cells, compositions comprising the engineered nucleic acids, the recombinant viruses, engineered cells, engineered proteins, chemical agents that are capable of activating expression of OCT4; KLF4; SOX2; or any combination thereof, an engineered protein selected from the group consisting of OCT4; KLF4; SOX2; or any combination thereof, an antibody capable of activating expression of OCT4; KLF4; SOX2; or any combination thereof, and methods of treating a (e.g., ocular disease), preventing a disease (e.g., ocular disease), regulating (e.g., inducing or inducing and then stopping) cellular reprogramming, regulating tissue repair, regulating tissue regeneration, or any combination thereof).

The invention discloses an mRNA-LNPs Brucella vaccine, an expression vector and a preparation method thereof. The mRNA-LNPs Brucella vaccine comprises at least one of mRNA of an outer membrane protein BP26 and mRNA of an outer membrane protein Omp 25. According to the present invention, the antigen sequence is optimized, the prepared mRNA Brucella vaccine can stimulate the body to produce strong immunogenicity, and compared with the Brucella vaccine prepared before the sequence optimization, the antibody titer is significantly improved. A mouse immune experiment research result shows that high-titer IgG, IgG1 and IgG2a can be generated. Th1 type reaction, Th2 type reaction or mixed Th type reaction is successfully induced.

The invention provides an alpha-glucosidase mutant and application thereof in synthesis of alpha-arbutin, and the alpha-glucosidase mutant comprises the following steps: maltose and hydroquinone are taken as substrates, the recombinant bacteria are taken as a catalyst, catalytic reaction is carried out under a heating condition, and the alpha-arbutin is obtained. According to the synthesis method, maltose and hydroquinone are used as reaction substrates, and alpha-arbutin is generated through whole-cell catalysis; meanwhile, the sequence of the alpha-glucosidase mutant is integrated onto a pRSEDuet-1 expression vector and transferred into escherichia coli BL21 (DE3) for expression, a reaction system of recombinant bacteria is optimized, the optimal catalytic condition is determined, the yield of alpha-arbutin under the optimal condition is 24.8 g / L, the yield can reach 95.6% and is improved by 1.2 times compared with that of an original strain, and the alpha-glucosidase mutant has a good application prospect.

The invention relates to the field of aquaculture, in particular to a preparation method and application of a larimichthys crocea iridovirus recombinant live vector vaccine. According to the invention, replication-deficient type 5 adenovirus is selected as a target gene expression vector, a recombinant adenovirus vector comprising main capsid protein of iridovirus is constructed, recombinant virus is obtained, and the safe and effective large yellow croaker iridovirus disease recombinant live vector vaccine is created. According to the vaccine, the larimichthys crocea is immunized in a soaking or bait feeding mode, the CPM of the larimichthys crocea immunized by the vaccine is 12.0%, and the CPM of the fish without being immunized by the vaccine is 62.0%. In addition, the RPS of the inoculated fish and the RPS of the empty carrier control group fish are 80.65% and 9.68% respectively. The method is at a leading level in the field of pseudosciaena crocea biomedicine at present, and creativity is remarkable.

The invention belongs to the technical field of plant heredity and genetic engineering, and particularly relates to a wheat saline-alkaline tolerant protein kinase gene TaCRCK and application thereof. Specifically, the invention discloses a wheat saline-alkaline tolerant protein kinase gene TaCRCK, and the nucleotide sequence of cDNA of the gene is as shown in SEQ ID NO. 1. The invention also discloses a plant expression vector containing the gene TaCRCK and application of the gene and the plant expression vector in cultivation of saline-alkali resistant plants. Tests prove that silent expression of the gene promotes the resistance of wheat to saline-alkaline stress, and the saline-alkaline tolerance of transgenic plants is obviously improved. The gene has a negative regulation effect on the salt and alkali tolerance of plants, provides a new tool and method for improving the salt tolerance of crops by using a genetic engineering means, and has important scientific significance and application prospects.