39 results about "Cloned genes" patented technology

The invention belongs to the technical field of gene engineering, and discloses a gene for regulating tillering, dry weight and yield of rice and application of the gene. The gene is an NIP1 gene and is located on the sixth chromosome of rice, the nucleotide sequence of the NIP1 gene is as shown in SEQ ID NO.1, and the amino acid sequence coded by the NIP1 gene is as shown in SEQ ID NO.2. The NIP1 gene is cloned, a transgenic vector is constructed, an NIP1 overexpression plant is obtained, and the NIP1 overexpression plant can regulate and control tillering, dry weight and yield of rice. Through determination of overexpression plant characters, compared with control group rice, overexpression shows tillering, dry weight and yield increase characters under low-nitrogen and high-nitrogen conditions, which indicates that the NIP1 gene plays an important regulation function in the aspects of tillering, dry weight, yield and nitrogen utilization of rice.

The application discloses application of a SdC2'H gene of Saposhnikovia divaricata and a method for increasing coumarin content in Saposhnikovia divaricata, and belongs to the technical field of bio-agriculture. The SdC2'H gene of Saposhnikovia divaricata and the biological material containing the SdC2'H gene are applied to the regulation of coumarin synthesis in Saposhnikovia divaricata. The method for increasing coumarin content in Saposhnikovia divaricata comprises the steps of increasing the expression amount and / or activity of the SdC2'H gene in Saposhnikovia divaricata. The SdC2'H gene is successfully cloned from Saposhnikovia divaricata for the first time, and a gene overexpression vector is constructed. The SdC2'H gene is positively regulated to control the synthesis of coumarin for the first time, and overexpression of the SdC2'H gene can significantly increase the product content. This not only deepens the understanding of the molecular mechanism of related biosynthesis, but also lays an important foundation for efficient production of high-value coumarin by using genetic engineering technology.

The invention discloses wheat TaPAB1 protein and application of related biological materials of the wheat TaPAB1 protein in the aspect of improving the grain weight of wheat. A gene for coding the TaPAB1 protein is named as a TaPAB1 gene. According to the invention, the gene TaPAB1 is cloned from a wheat cultivation variety, Chinese spring, and is over-expressed in a wheat variety Kenong 199 (KN199) by utilizing a gene engineering technology, so that the thousand seed weight, the grain length, the plant height, the ear length and the yield of a TaPAB1 over-expressed plant are obviously increased compared with those of a wild type. The invention provides a research basis for increasing the wheat yield and breeding high-yield varieties.

The invention belongs to the field of plant genetic engineering and discloses a cold-resistant gene of wild grape VaWRKY65 Application in genetic improvement of plant cold resistance, VaWRKY65 The gene is from a very cold-resistant wild grape ( Vitis amurensis ) was isolated and cloned, and its sequence is shown in SEQ ID NO. 1. This gene was used to construct overexpression and interference vectors, and was introduced into Arabidopsis thaliana and Vitis amurensis respectively through Agrobacterium-mediated genetic transformation. The obtained transgenic plants were verified by biological function, indicating that the cloned gene of the present invention VaWRKY65 The gene has the function of improving plant cold resistance. The discovery of this gene provides a new genetic resource for molecular design and breeding of plant stress resistance, and provides a new genetic resource for the implementation of green agriculture and water-saving agriculture. The development and utilization of this genetic resource will help reduce agricultural production costs and achieve environmental friendliness.

This invention discloses the application of the CLE11 small peptide gene in regulating plant salt tolerance, belonging to the field of biomaterials technology. This invention uses millet as the research material and clones... SiPROCLE11 Gene (CLE11 small peptide gene), construction SiPROCLE11 Overexpression and silencing of millet materials, exogenous application of the synthetic peptide SiCLE11p, and its role in enhancing millet salt tolerance were determined. Further investigation revealed that it can promote lateral root development and reduce Na+ in the roots. + This accumulation of knowledge laid the foundation for the discovery of salt-tolerant genes in millet and for salt-tolerant millet breeding.

The application discloses C4 corn ZmClpP6 protein and application of related biological materials thereof in regulation of high light response capability of plants. ZmClpP6 It is found that: ZmClpP6 The knockout can reduce chlorophyll content, total carotenoid content, catalase activity and copper-zinc SOD enzyme activity of corn under high light conditions, and makes corn leaves appear yellowing phenotype. It is indicated that ZmClpP6 can regulate high light response capability of plants. The application clones ZmClpP6 the gene in corn for the first time and analyzes the biological function thereof, and has important significance for cultivation of high light efficiency plants.

The present invention belongs to the field of biological genetic engineering and relates to the use of ginseng transcription factor PgERF2 in improving plant resistance to Alternaria alternata. The sequence of the ginseng transcription factor PgERF2 protein is shown in SEQ ID NO.1; the sequence of the ginseng transcription factor PgERF2 gene is shown in SEQ ID NO.2. The present invention clones the PgERF2 gene and constructs a corresponding plant expression vector, successfully transferring the gene into wild-type Arabidopsis thaliana. Transcriptomic analysis results show that the response to biotic stress is activated in the transgenic plants, and the expression level of the plant defense protein-related gene AtPDF1.2A is significantly increased. It is also demonstrated that PgERF2 can directly bind to the GCC-box element to activate the expression of the plant defense protein-related gene AtPDF1.2A, thereby significantly enhancing resistance to Alternaria alternata.

The invention belongs to the technical field of gene engineering, and discloses a gene for regulating and controlling the nitrogen utilization rate of rice and application of the gene. The gene is OsDLN13, the nucleotide sequence of the gene is shown as SEQ ID NO.1, and the gene is located on a first chromosome of rice. The gene OsDLN13 can regulate and control the plant height, tillering, single plant yield and nitrogen absorption capacity of rice. By cloning the gene OsDLN13, constructing a transgenic vector, obtaining an overexpressed plant and measuring the phenotypic character of the transgenic plant, it is found that the plant height, the tiller number, the yield of a single plant and the nitrogen absorption capacity of the overexpressed plant are remarkably changed, and compared with wild type rice, the plant height, the tiller number, the yield of a single plant and the nitrogen absorption capacity of the overexpressed plant are remarkably improved. The overexpressed plant shows the characters of plant height, tillering number, single plant yield, nitrogen absorption capacity increase and the like, which indicates that the OsDLN13 gene plays an important regulation and control role in rice plant height, tillering number, single plant yield and nitrogen absorption capacity.

This invention provides rice OsPDRA3 Application of genes in improving insect resistance in rice. This invention clones genes from the rice genome. OsPDRA3 Gene (LOC_Os01g42380), analyzed using the SMART program, predicts that this protein possesses multiple transmembrane domains and ATPase domains (AAA+) associated with various cellular activities. Further investigation is needed. OsPDRA3 The effect on tolerance of rice brown planthopper was investigated by comparing wild-type and overexpression. OsPDRA3 The tolerance of transgenic plants to brown planthoppers was assessed, and the results showed that overexpression of [the planthopper species]... OsPDRA3 Compared to wild-type rice lines (0% survival rate), the survival rate of rice lines treated with brown planthoppers was significantly improved, reaching 75%–100%. This indicates that… OsPDRA3 Genes can significantly improve the insect resistance of rice and can be widely used in the fields of plant genetic engineering and plant disease resistance, with huge economic value and application prospects.

The invention relates to application of a brassica napus BnaA08. SNAT2 gene in improving the seed germination rate under drought stress, in particular to a method for improving the seed germination rate of brassica napus under drought stress. According to the invention, a BnaA08. SNAT2 gene is cloned from a brassica napus inbred line 'K407', an overexpression vector is constructed, and a transgenic line is obtained through agrobacterium transformation. Functional verification shows that the germination potential and germination rate of a transgenic line under drought stress (simulated by D-mannitol) are remarkably higher than those of a wild type, the content of hydrogen peroxide and superoxide anions is reduced, and the activity of antioxidant enzyme and the content of proline are increased. According to the breeding method provided by the invention, the drought tolerance of the plant seeds in the germination period can be improved by increasing the content and / or activity of the BnaA08. SNAT2 gene coding protein in the plant.

The present invention belongs to the field of biological genetic engineering and relates to the application of the ginseng transcription factor PgJAZ13 in enhancing tobacco brown spot disease resistance. The sequence of the PgJAZ13 gene is shown in SEQ ID NO.2. The present invention clones the PgJAZ13 gene and constructs a corresponding plant expression vector. Through an Agrobacterium-mediated transformation method, the gene is successfully transferred into wild-type tobacco leaves, and positive tobacco plants that stably overexpress the PgJAZ13 gene are screened. Experiments have shown that heterologous overexpression of the PgJAZ13 gene in tobacco can significantly increase the expression levels of genes related to disease resistance and key enzyme genes for terpenoid synthesis, thereby promoting the biosynthesis of disease-resistant proteins and terpenes, and ultimately enhancing the plant's disease resistance.

The application belongs to the field of biological genetic engineering and relates to application of PgMYC3 in regulation of ginsenoside biosynthesis. The sequence of the ginseng transcription factor PgMYC3 is shown as SEQ ID NO. 2. The application clones the PgMYC3 gene, constructs a plant expression vector, and adopts the Agrobacterium-mediated transformation method to transform ginseng callus. Through RT-qPCR technology identification, positive callus overexpressing PgMYC3 is obtained. In the transgenic ginseng callus, the total saponin content is increased by 1.5 times, which indicates that PgMYC3 has a promoting effect on the accumulation of ginsenosides. Meanwhile, through molecular biology technology, it is confirmed that PgMYC3 can directly bind to the G-box element to activate the expression of the saponin synthesis gene PgPPDS.

The invention discloses application of a CLE11 small peptide gene in regulation and control of plant salt tolerance, and belongs to the technical field of biological materials. According to the invention, foxtail millet is used as a research material, a SiPROCLE11 gene (CLE11 small peptide gene) is cloned, a SiPROCLE11 overexpression and silent foxtail millet material is constructed, an artificially synthesized small peptide SiCLE11p is externally applied, and the effect of the small peptide SiCLE11p on enhancing the salt tolerance of foxtail millet is determined. Furthermore, it is found that the Setaria italica can promote lateral root development of Setaria italica and reduce Na < + > accumulation in root systems, and a foundation is laid for digging of salt-tolerant related genes of Setaria italica and salt-tolerant breeding of

The application belongs to the technical field of genetic engineering, and particularly relates to a CsECT protein related to the disease resistance of plants, a recombinant vector and application thereof. 6 The CsECT protein provided by the application is synthesized by the gene CsECT, and the gene CsECT is speculated to be an m A reader, which affects the metabolism of target mRNAs, improves the stability of disease resistance related target genes, promotes the expression of disease resistance genes, and further improves the disease resistance of plants. The results of the examples show that the CsECT gene is cloned to construct a CsECT transgenic citrus, the transgenic citrus overexpresses the CsECT gene, and the disease resistance of the citrus to bacterial wilt can be significantly improved, thereby improving the disease resistance of the citrus. It can be seen that the CsECT protein synthesized by the gene CsECT can improve the disease resistance of plants.

This invention provides GhFRC1 The application of genes in regulating plant fertility belongs to the field of plant genetic engineering technology. This invention is the first to clone genes from cotton. GhFRC1 The gene was identified and verified to be heterologously expressed in three different genera and families of crops: Arabidopsis thaliana, tomato, and rice, and to play a role in fertility regulation. This breakthrough overcomes the limitation of existing fertility genes that can only regulate the fertility of a single crop, and realizes "broad-spectrum regulation of fertility of multiple crops by a single gene", filling the gap in the application of distant heterologous expression of cotton fertility genes. GhFRC1 After heterologous expression of the gene, it can stably lead to inactive pollen and abnormal fruit set in the recipient crop, and can also cause a phenotype of complete pollen sac non-development. Moreover, the sterility trait can be restored to fertility through normal pollen pollination. This solves the problem of the narrow applicability of existing fertility genes and the great limitation of their breeding applications, and provides new gene resources and technical solutions for fertility regulation of multiple crops and hybridization breeding.

The present application relates to the field of agricultural biotechnology, in particular to locust immune related protein LmEaster gene and application thereof. The present application clones LmEaster gene and synthesizes double-stranded RNA, adopts injection method to treat locusts, and detects the mortality of locusts in each treatment and the expression amount change of related antibacterial peptide, and confirms that after interfering with LmEaster gene of locusts, the expression amount of antibacterial peptide of locusts is increased, the immune defense ability of locusts is increased, and then the mortality is reduced.

The invention relates to the technical field of plant molecular biology, in particular to application of a Unicaulis gene for regulating and controlling growth of tomato axillary buds in tomato breeding. The Uniicaures gene is used for positively regulating and controlling the elongation and growth of lateral branches of tomatoes, and the nucleotide sequence of the Uniicaures gene is as shown in SEQ ID NO. 1; the amino acid sequence of the encoding protein of the Unicaulis gene is as shown in SEQ ID NO. 2. The tomato lateral branch elongation growth gene Unicaulis is positively regulated and controlled by constructing a genetic linkage group, finely positioning and cloning, and by overexpressing the Unicaulis gene in a tomato lateral branch-free plant or increasing the content of the encoding protein of the Unicaulis gene, the accumulation amount of auxin at the axilla part of a transgenic plant is reduced, and the lateral branch of the tomato can normally sprout and obviously grow. The Unicaulis gene or the encoded protein thereof provided by the invention can significantly reduce the accumulation amount of auxin in the axillary part of the tomato plant, relieve the apical dominance of the tomato plant to lateral branches, accelerate the extension and growth of axillary buds of the axillary part of the tomato plant into lateral branches, and provide gene resources and new germplasm for cultivating tomato varieties with strong lateral branches.

The invention discloses application of a radish RsBBX9 gene in regulation and control of anthocyanin synthesis and a method, and belongs to the technical field of plant genetic engineering. According to the invention, an RsBBX9 gene is cloned, an overexpression vector pFGC1008-RsBBX9 and a VIGS silence vector pTY-S-RsBBX9 are constructed, and transient expression and gene silencing technologies are combined, so that the regulation and control effect of the RsBBX9 on the anthocyanin content is verified. Yeast two-hybridization, BiFC and pull-down experiments further prove that RsBBX9 and RsMYB1 interact to synergistically activate an anthocyanin synthesis pathway. The invention provides an efficient technical scheme for radish variety improvement and anthocyanin production.

The invention discloses application of a radix saposhnikoviae SdC2 'H gene and a method for increasing the content of coumarin in radix saposhnikoviae by using the SdC2' H gene, and belongs to the technical field of biological agriculture. The divaricate saposhnikovia root SdC2 'H gene and the biological material containing the divaricate saposhnikovia root SdC2' H gene disclosed by the invention are applied to regulation and control of synthesis of divaricate saposhnikovia root coumarin. The method for increasing the content of coumarin in radix saposhnikoviae comprises the step of increasing the expression quantity and / or activity of an SdC2 'H gene in radix saposhnikoviae. The SdC2 'H gene is successfully cloned from radix saposhnikoviae for the first time, and a gene overexpression vector of the SdC2' H gene is constructed. The key function of the SdC2 'H gene for positively regulating the synthesis of coumarin is clearly disclosed for the first time, and the product content can be remarkably increased through overexpression of the SdC2' H gene, so that the understanding on a related biosynthesis molecular mechanism is deepened, and an important foundation is laid for efficiently producing high-value coumarin by utilizing a gene engineering technology.

The application discloses a citrus transcription factor gene and application thereof in promoting fruit ripening CsAP2-16 . The gene belongs to an AP2 / ERF family member, the CDS sequence of which is shown as SEQ ID NO. 1, the CDS sequence length is 1290 bp, 429 amino acids are encoded, and the amino acid encoded is shown as SEQ ID NO. 2. A primer is designed to amplify the gene from Fengjie navel orange CsAP2-16 . The gene is introduced into Mic-Tom tomato and golden orange by using an agrobacterium-mediated genetic transformation method, and the obtained transgenic plants are verified by phenotype observation statistics and gene expression analysis, and it is shown that the cloned gene has the function of significantly promoting citrus fruit ripening. CsAP2- 16 The application provides a new gene resource for molecular design breeding of fruit ripening.

The application discloses a method for analyzing epigenetic diversity based on a methylation-sensitive amplified polymorphism (MSAP) gene and a biological gene fingerprint table. The method comprises the following steps: obtaining an MSAP amplification electropherogram of a biological sample to be detected, and constructing a 01 matrix of the sample; converting the 01 matrix into an MSAP gene sequence by using a "CCGGN" replacement method; comparing the MSAP gene sequence with a reference genome, and annotating DNA methylation related gene information; based on the annotation result, constructing a gene fingerprint table based on the copy number of the methylation gene and the methylation / hemimethylation mode, and analyzing epigenetic diversity. The method can convert a traditional MSAP band fingerprint map into a gene fingerprint table with annotation function information without performing cloning and sequencing of MSAP differential fragments, can improve MSAP marker analysis to epigenome analysis, solves the problem that MSAP epigenes are difficult to clone, and improves the level of germplasm resource identification and molecular breeding.

The invention relates to the technical field of biology, in particular to application of ZmATG13a protein and a coding gene thereof to regulation and control of corn yield, ZmATG13a gene is cloned in corn, CRISPR / Cas9 mutant plants of corn ZmATG13a are obtained through genetic transformation, corn selfing line groups are sown in a sufficient phosphorus land parcel, and it is found that the zmatg13a mutant plants have higher plant height than wild type spinning periods, so that the ZmATG13a protein can be used for regulating and controlling corn yield. The stem diameter is not obviously different from that of a wild type; the ear length of the zmatg13a mutant plant is obviously longer than that of a wild type plant, and the single ear weight, the ear grain weight and the hundred-grain weight of the zmatg13a mutant plant are all larger than those of the wild type plant, so that the zmatg13a mutant plant shows the advantage in yield. The ZmATG13a is a protein with an ATG13 structural domain, and the ZmATG13 gene is knocked out through a gene editing method, so that the plant height and the yield in the spinning period are remarkably increased, and a germplasm resource is provided for cultivating a new variety.

The invention provides application of a rice OsPDRA3 gene in improving insect resistance of rice. According to the invention, an OsPDRA3 gene (LOCOs01g42380) is cloned in a rice genome, and an SMART program is used for analyzing and predicting that the protein has multiple transmembrane domains and ATP enzyme structural domains (AAA < + >) related to various cell activities. The influence of OsPDRA3 on the tolerance of rice brown planthopper is further studied, and by comparing the tolerance of wild type and overexpressed OsPDRA3 transgenic plants to brown planthopper, the result shows that the survival rate of a rice line with overexpressed OsPDRA3 after being treated by brown planthopper is obviously increased compared with that of a wild type rice line (the survival rate is 0%), and reaches 75%-100%. Namely, it is shown that the OsPDRA3 gene can significantly improve the insect resistance of rice, can be widely applied to the fields of plant genetic engineering and plant disease resistance, and has huge economic value and application prospects.

The invention relates to the field of gene breeding, in particular to application of FveJAZ5 protein and a coding gene thereof in regulation and control of plant anthracnose resistance. According to the invention, the FveJAZ5 gene is cloned in strawberry, and a strawberry FveJAZ5 overexpression plant is obtained through genetic transformation. In vitro leaves of a wild type, an FveJAZ5 overexpressed leaf and an FveJAZ5 gene editing mutant are subjected to colletotrichum inoculation treatment respectively, culture and phenotype observation are carried out under the conditions of 28 DEG C and 100% relative humidity, a scab area statistical result shows that the scab area of the FveJAZ5 overexpressed leaf is remarkably smaller than that of the wild type, and the scab area of the FveJAZ5 gene editing mutant leaf is remarkably larger than that of the wild type; it is indicated that the anthracnose resistance of the FveJAZ5 overexpression plant is remarkably improved, and the whole growth of the FveJAZ5 overexpression plant has no adverse phenotype compared with that of a wild type, so that the FveJAZ5 plays an important role in the strawberry anthracnose resistance regulation and control process, and the breeding potential is good.

The application discloses a wheat disease resistance related protein TaNLR-4B and a coding gene and application thereof, and belongs to the technical field of genetic engineering. The amino acid sequence of the TaNLR-4B provided by the application is shown as SEQ ID NO:1, the ORF sequence of the TaNLR-4B is shown as SEQ ID NO:2, the gene TaNLR-4B is induced to express in a wheat stripe rust compatible interaction system, is not induced to express in a wheat stripe rust non-compatible interaction system, and plays a positive regulation role in the compatible interaction of the wheat and the stripe rust fungus. The gene TaNLR-4B is cloned in Gui Xie 3, overexpressed in a wheat stripe rust susceptible variety Fielder, TaNLR-4B overexpression plants are obtained, and the enhanced resistance of the TaNLR-4B overexpression plants to the stripe rust fungus CYR34 is verified, thereby providing a new gene resource for wheat stripe rust resistance breeding.

The invention belongs to the technical field of plant genes, and particularly relates to application of TabZIP4 to improvement of drought resistance of wheat. The base sequence of the TabZIP4 is shown as SEQ ID NO.9, and the amino acid sequence of the TabZIP4 is shown as SEQ ID NO.10; according to the application, a knockout strain is prepared by knocking out TabZIP4 or reducing the expression quantity of TabZIP4, so that the drought resistance of wheat is improved. The knockout strain of the TabZIP4 is obtained by utilizing a transgenosis method, the function of the knockout strain is deeply analyzed and identified, and the effect of the TabZIP4 gene in the process that the wheat responds to the drought stress is further clarified. The cloned TabZIP4 gene provides a new wheat drought-resistant related gene, and provides a new clue for wheat drought-resistant breeding.

A method of erasing an abnormal epigenetic modification in a sheep iCHI embryo and use thereof. A method including the steps of: transferring a recombinant plasmid expressing protamine into a sheep androgenetic haploid embryonic stem cell, then introducing the cell into a mature sheep oocyte, and performing activation and culture to obtain a sheep iCHI embryo without abnormal epigenetic modifications. In the present disclosure, abnormal methylations in iCHI embryos may be erased by transient expression of protamine in o-haSCs. The blastocyst rate of the prepared Pro-iCHI embryos is significantly higher than that of iCHI embryos, and is similar to that of IVF embryos. The resulting embryos can be used for embryo transfer to obtain semi-cloned gene-edited sheep, providing materials for subsequent breeding applications.

The invention belongs to the technical field of plant molecular breeding, and particularly relates to a gene for ubiquitination regulation and control of flowering of eucommia ulmoides and application of the gene. A yeast two-hybrid technology is used for screening interaction protein of the eucommia ulmoides flowering inhibitor gene FLC, a full-length transcriptome database of eucommia ulmoides is annotated as ubiquitination E3 binding enzyme, the encoding gene of the EuUBL2 is named as EuUBL2, the EuUBL2 gene is cloned from eucommia ulmoides through an RACE technology, the gene sequence and translated amino acid of the EuUBL2 gene are shown in a figure 2, the full length of cDNA of the EuUBL2 gene is 1175 bp, the ORF (Open Reading Frame) (ORF, 49-1063 bp) is 1014 bp, and the ORF (Open Reading Frame) is 496bp. And a protein containing 337 amino acid residues is coded. According to the application, the EuUBL2 gene is over-expressed, and the plant has an early flowering phenotype. The result shows that EuUBL2 ubiquitination EuFLC regulates and controls early blossoming of arabidopsis thaliana. The method is further applied to plant breeding by utilizing a bioengineering technology, and a research basis can be provided for cultivating a new variety of plant early blossoming.

The application discloses a cold-resistant gene CmMYB106 of chrysanthemum, wherein the nucleotide sequence is shown as SEQ ID NO:1 in a sequence listing, and the application also discloses a primer pair CmMYB106-qF and CmMYB106-qR for cloning the cold-resistant gene CmMYB106, wherein the nucleotide sequences are shown as SEQ ID NO:2 and SEQ ID NO:3 in the sequence listing. The application takes chrysanthemum as a material, clones the CmMYB106 gene, and determines the expression characteristics of the CmMYB106 gene by means of expression modes and transcription activation activities. An overexpression vector is constructed, and the chrysanthemum is genetically transformed by means of agrobacterium, and the influence of the CmMYB106 on the cold resistance of chrysanthemum is analyzed, thereby providing a theoretical basis for chrysanthemum cold resistance breeding.

The present invention discloses the rational transformation of L-threonine transaldolase and its integrated system for the efficient synthesis of β-hydroxy-α-amino acids, belonging to the field of biocatalytic engineering. The present invention clones the gene of L-threonine transaldolase from Burkholderia diffusa and heterologously expresses it in E.coli BL21 (DE3) to study the enzymatic properties of its recombinant enzyme and the function of catalytic conversion of L-threo-methylsulfonylphenylserine. At the same time, the key amino acid sites of L-threonine transaldolase are mutated to obtain mutants with significantly improved catalytic efficiency and stereoselectivity. The present invention also introduces an L-threonine transaldolase-mediated catalytic system into an efficient by-product acetaldehyde elimination system to construct an integrated system; not only the inhibitory effect of by-products on the catalytic reaction is eliminated, but also the solubility of the substrate is improved by adding the auxiliary substrate isopropanol, while achieving the in situ regeneration of the cofactor NADH required for the reaction.