865 results about "Key genes" patented technology

The invention belongs to the technical field of biology, and relates to an escherichia coli engineering strain for producing L-isoleucine through a methyl malic acid way as well as a construction method and application of the escherichia coli engineering strain. According to the invention, Escherichia coli BW25113 is used as a chassis strain, and a methyl malic acid synthase high-activity mutant gene cimA3.7, an isopropyl malic acid isomerase gene GsleuCD, a 3-isopropyl malic acid dehydrogenase gene AfleuB and a leucine dehydrogenase gene Lsleudh are over-expressed; a methyl malic acid absorption protein gene is over-expressed, and a branched chain amino acid transport system is modified to improve the exosome of the L-isoleucine; all methyl malic acid pathway related genes are integrated to a genome of a chassis strain, and a plasmid-free L-isoleucine production strain is constructed; the copy number of a key gene on a genome is optimized, and the yield of the engineering strain L-isoleucine is increased by introducing a non-oxidative glycolysis approach. The yield of the L-isoleucine after the engineering strain is fermented for 34 hours in a fermentation tank is 56.6 g / L.

The invention discloses a sugarcane germplasm resource evaluation and breeding method for smart agriculture. The sugarcane germplasm resource evaluation and breeding method comprises the following steps: step 1, collecting materials with wide genetic diversity; establishing an intelligent incubator and a greenhouse, and monitoring environmental parameters by using an Internet of Things sensor; 2, performing data acquisition on the related germplasm resources by using an unmanned aerial vehicle, a robot and a near infrared spectrum technology, and constructing a phenotype database for data management; meanwhile, genotype identification is carried out by combining molecular marker-assisted selection and genome selection technologies, molecular markers of key genes are mined, and the genetic value of related germplasm is evaluated; step 3, constructing a hybrid combination prediction model by using an AI algorithm, optimizing parent matching of the related germplasm and the cultivated sugarcane, monitoring and collecting the filial generation at the same time, and analyzing and screening in combination with smart agriculture; 4, demonstration planting is conducted on the bred excellent sugarcane strain, and application of the excellent strain is improved through government-enterprise-scientific research institution collaborative popularization and in combination with farmer technical training.

The invention discloses a biosynthesis system for high-yield 1, 6-hexamethylenediamine and application of the biosynthesis system, and belongs to the technical field of biology. According to the method, key genes which are excavated and optimized in a 1, 6-hexamethylenediamine biosynthetic pathway are co-expressed in a single cell module or a multi-cell module, and different enzyme elements expressed by the cell module are subjected to enzyme cascade reaction, so that 1, 6-hexamethylenediamine can be efficiently synthesized by taking caprolactam or 6-aminocaproic acid as a substrate; furthermore, after the carboxylic acid reductase MaCAR is mutated, the catalytic activity of the carboxylic acid reductase MaCAR can be remarkably improved, and the yield of the 1, 6-hexamethylenediamine is further improved; in addition, the biosynthesis system disclosed by the invention directly takes the cell module as a whole-cell catalyst, and an enzyme purification step is avoided, so that the biosynthesis system has the advantage of low cost.

The invention is applicable to the technical field of bioengineering, and provides a coding gene for enhancing iron deficiency stress tolerance of plants and application. According to the soybean GmMYB14 transcription factor disclosed by the invention, the problem that the growth of soybeans is limited in an iron-deficient environment, particularly calcareous soil and soda saline alkali soil, is solved by excavating and applying the encoding gene (the nucleotide sequence is shown as SEQ ID No.1) of the soybean GmMYB14 transcription factor. Experiments prove that after the gene is over-expressed in arabidopsis thaliana and soybean, the tolerance of plants to iron deficiency stress can be remarkably enhanced. The invention provides a key gene resource for molecular breeding: the GmMYB14 effectively promotes the absorption and utilization of plants on iron by regulating and controlling the metabolic pathway of phenylpropane, not only can improve the planting adaptability of soybeans in low-iron soil, but also can improve the yield and quality of the soybeans, provides powerful technical support for enlarging the planting area of the soybeans in iron-deficient soil regions, and has a wide application prospect. The method is of great significance in relieving insufficient soybean productivity in China and improving the self-sufficiency rate.

The invention relates to a mulberry MaBEH1 / 2 gene for improving plant salt tolerance and application thereof, and belongs to the technical field of plant genetic engineering and biology. Wherein the coding nucleotide sequence of the mulberry MaBEH1 / 2 gene for regulating and controlling the salt tolerance of the woody plant is shown as a sequence 3; and the amino acid sequence of the MaBEH1 / 2 expression protein is shown as a sequence 4. According to the invention, MaBEH1 / 2 is transferred into 84K poplars to obtain MaBEH1 / 2-OE transgenic poplars; compared with a non-transgenic poplar, the salt stress tolerance of the MaBEH1 / 2-OE transgenic poplar can be remarkably enhanced. The invention provides key gene resources and technical support for genetic improvement of salt-tolerant forest varieties.

The invention discloses development and application of a molecular marker of a rape stain tolerance related gene BnaERF3, and belongs to the technical field of molecular biology. According to the invention, the BnaA06. ERF3 gene is screened on the basis of whole genome associated molecules, and is used as a key gene in QTL (qWL.A06) to participate in regulation and control of the waterlogging tolerance of the rape. A molecular marker related to the waterlogging tolerance of the brassica napus is developed aiming at the BnaA06. ERF3 gene, and tests show that the waterlogging tolerance of the brassica napus can be detected by utilizing the molecular marker, so that varieties containing waterlogging tolerance genes are subjected to auxiliary selection and breeding, and the waterlogging tolerance breeding process of the brassica napus is accelerated.

The invention discloses a key gene identification method related to tobacco nitrogen response, which comprises the following steps: S1, setting four nitrogen fertilizer gradient treatments on the basis of same phosphorus and potassium fertilization by adopting a field experiment of a completely random block; s2, randomly taking the 6th to 8th leaves of the five plants in each area, and dividing a sample into two parts: quickly freezing one part with liquid nitrogen, and storing at-80 DEG C for RNA (Ribonucleic Acid) extraction and transcriptome sequencing; one part is used for measuring the nitrogen content, and after baking and drying treatment, a KjeltecTM8100 automatic nitrogen analyzer is used for measuring; s3, nitrogen content determination: determining the nitrogen content in the treated sample by using an automatic nitrogen analyzer KjeltecTM8100; according to the method, high-throughput transcriptome sequencing and weighted gene co-expression network analysis (WGCNA) are combined, so that not only can gene expression maps of flue-cured tobacco leaves treated by different nitrogen fertilizers be comprehensively captured, but also gene modules with similar expression modes can be mined from a global perspective.

The invention discloses a rice drought resistance phenotype detection and genetic analysis method based on a phenotypic robot, which is characterized in that rice hyperspectral phenotypic character data is acquired based on the phenotypic robot, and the relative water content RWC of rice is predicted. Through high-frequency data acquisition, RWC (t) based on a time sequence is obtained, RWC (t) curve change characteristic indexes are calculated, the dynamic change process of a rice experiment material in the whole drought experiment period is quantified, the drought resistance index DTI of the rice experiment material is obtained, and the drought resistance of the rice variety is comprehensively evaluated. DTI as a user-defined phenotypic character can be used for whole genome association analysis (GWAS), and a key gene for controlling the drought resistance of rice is excavated. According to the invention, the problems of low efficiency and low flux of traditional rice phenotype detection are solved, phenotype-gene mining based on the mobile robot is realized, and a new way is expanded for improvement of rice genetic breeding.

The invention discloses a GmbZIP36 protein related to aluminum resistance of plants in acid soil as well as a coding gene and application of the GmbZIP36 protein, and belongs to the technical field of bioengineering. The amino acid sequence of the GmbZIP36 protein is shown as SEQ ID NO: 2, 3 or 4, and the nucleotide sequence of the coding gene of the GmbZIP36 protein is shown as SEQ ID NO: 1. The invention finds that the expression of the GmbZIP36 gene is induced by aluminum stress, the relative root elongation and root biomass of arabidopsis thaliana and soybean under the aluminum stress can be obviously improved by over-expressing the gene in a plant, and aluminum accumulation is reduced, so that the aluminum resistance of the plant is enhanced. The invention provides a recombinant vector containing the gene, a host cell and application of the recombinant vector and the host cell in regulating and controlling the aluminum resistance of plants. Meanwhile, the invention also provides a method for producing an aluminum-resistant transgenic plant by using the gene, a kit for detecting the gene or protein and a method for carrying out assisted breeding by using the molecular marker linked with the gene. The invention provides a key gene resource for aluminum-resistant molecular breeding of crops, and has important significance for cultivating new varieties of crops adapting to acid soil.

The invention relates to the technical field of tumor data processing, in particular to a methylation-based tumor data processing system. The system comprises the following modules: a methylated sample sequencing module, a sequencing difference site recognition module, a tumor gene sequence analysis module and a tumor subtype classification module, performing high-throughput sequencing on the to-be-detected clinical tumor DNA sample to obtain methylated tumor sequencing data; performing methylation level quantification on the methylated tumor sequencing data, and performing tumor difference site analysis to generate tumor difference site data; performing tumor generation key gene sequence identification according to the tumor difference site data to generate tumor methylation characteristic data; and performing tumor subtype tag identification according to the tumor methylation characteristic data to generate tumor type tag data. According to the method, accurate tumor typing is realized on the basis of tumor DNA methylation characteristic analysis, and the false negative rate of low-concentration sample detection is effectively reduced.

The invention discloses a pichia pastoris engineering bacterium for synthesizing bakuchiol from de novo by converting methanol as well as construction and application of the pichia pastoris engineering bacterium. A heterologous p-coumaric acid synthesis pathway is introduced to neutral sites of different chromosomes of pichia pastoris, a high-yield p-coumaric acid strain is obtained through gene knockout, overexpression or heterologous expression of tyrosine and phenylalanine synthesis pathway key genes of a shikimic acid pathway, a heterologous bakuchiol biosynthesis pathway is introduced on the basis, and a high-yield p-coumaric acid strain is obtained. After overexpression of bakuchiol synthetase, endogenous overexpression or heterologous expression of MVA pathway key genes and exogenous acetyl coenzyme A supply pathway genes, methanol concentration optimization and bacterial strain His4 gene supplementation, the yield of bakuchiol is effectively increased to 91.2 mg / L and is increased by 59.8 times compared with that of an initial bacterial strain, and the yield of bakuchiol in a 15L fermentation tank reaches 692.8 mg / L. The method has the characteristics of high conversion efficiency, low production cost, convenience in preparation, wide industrial application prospect and the like.

The invention relates to the technical field of cotton alkali-resistant gene mining, and particularly discloses a method for jointly screening key genes responding to upland cotton alkali stress based on transcriptome and metabolome, which comprises the following steps: carrying out alkali stress treatment on upland cotton to obtain tissue samples at different time points; performing transcriptome sequencing and metabolome sequencing on the tissue sample to respectively obtain gene expression data and metabolite data; performing difference analysis on the gene expression data and the metabolite data to obtain DEGs and DAMs; carrying out conjoint analysis on the DEGs and the DAMs, and screening to obtain a key gene and a regulatory pathway which respond to alkali stress; analyzing the key gene and the regulatory pathway through a gene expression verification technology to obtain an expression mode; according to the invention, 15 candidate genes related to the alkali resistance of the upland cotton are screened through WGCNA, a theoretical basis is provided for deeply knowing the molecular mechanism of the alkali resistance of the upland cotton, and a new gene resource is provided for subsequent research.

The invention relates to cloning of a stress-resistant gene LpbHLH75 of Lilium pumilum DC (Lilium pumilum DC.) and application of the stress-resistant gene LpbHLH75 of Lilium pumilum DC. The lilium tenuifolium stress resistance key genes are mined through transcriptome data analysis, and the functions of the lilium tenuifolium stress resistance key genes are studied. Comprising the following steps: (1) cloning a target gene; (2) constructing a plant overexpression vector; and (3) detecting physiological indexes of the transgenic plant. According to the experiment, lilium tenuifolium is taken as a research object, and the LpbHLH75 gene is cloned by applying an RT-PCR technology; transforming tobacco by constructing an overexpression vector; the function of overexpression of the LpbHLH75 on tobacco salt and alkali and drought stress is identified through transgenic tobacco, the function of the lilium tenuifolium bHLH gene under adversity stress is disclosed, a new gene source is provided for lilium tenuifolium genetic engineering breeding, a reference is provided for later lilium tenuifolium stress-resistant germplasm resource research, and the lilium tenuifolium bHLH gene is developed. And a foundation is laid for exploring a gene capable of enhancing stress resistance in the lilium tenuifolium and a stress resistance mechanism of the gene.

The invention relates to the technical field of agricultural biology, and particularly provides a dsRNA nanocrystallization preparation for preventing and treating areca yellows as well as a preparation method and application of the dsRNA nanocrystallization preparation. Areca yellows are a destructive disease caused by areca symptomless virus type 1 (APV1), eight key target gene segments of an APV1 virus genome are screened, specific double-stranded RNA is designed, a nano-carrier is used for wrapping and delivering a dsRNA mixture obtained by transcription of the eight target gene segments, and the areca yellows are obtained by screening. And a nanocrystallization preparation for improving the environmental stability and RNA interference efficiency of the dsRNA is prepared. Experimental results show that the nanocrystallization preparation can effectively inhibit the key gene expression of APV1 virus, reduce the virus load and improve the resistance of areca catechu to yellows. The invention provides an accurate, efficient and environment-friendly biological prevention and control strategy, and has a wide agricultural application prospect.

The invention relates to a key gene GqBurscon for wing development of a Qinghai grassland moth and application thereof. The key gene GqBurscon for the wing development of the Qinghai grassland caterpillar has a nucleotide sequence as shown in SEQ ID NO: 1. According to the dsRNA synthesized by the key gene GqBurscon for wing development of the Qinghai grass caterpillar, one chain of the dsRNA is the same as the sequence of SEQ ID NO: 1, and the other chain of the dsRNA is reversely complementary with the sequence of SEQ ID NO: 1. The key gene GqBurscon for the wing development of the Qinghai grassland is used for synthesizing dsRNA, the synthesized dsRNA is used for carrying out RNA interference on a target gene, the gene transcription level is knocked down, the gene is effectively silenced in chest, abdomen, wings and wing base tissues of male adult Qinghai grassland, the length and width of the wings are correspondingly reduced, the development of the wings is limited, and the gene transcription level is reduced. The flying capability is influenced, and the death is caused.

The invention discloses a TaIMPA1 gene and application of the TaIMPA1 gene in improvement of plant genetic transformation efficiency. The nucleotide sequence of the gene is shown as SEQ ID No: 1 in a sequence table. According to the invention, the key gene TaIMPA1 is cloned from a plant body to delivery for the first time, and the genetic transformation performance is enhanced by remarkably improving the delivery efficiency of exogenous DNA (Deoxyribose Nucleic Acid). The discovery fills up the research blank of plant endogenous delivery regulatory factors, and an original solution is provided for overcoming the core problem of genotype dependence.

The invention discloses a neural network calculation method and device for gene expression regulation and control analysis, and relates to the technical field of bioinformatics, and the method comprises the steps: obtaining first feature data and second feature data; constructing an input feature comprising a plurality of regulation and control hierarchies; and inputting the input features of the plurality of regulation levels and the second feature data into the target neural network model, and outputting a prediction result of the gene expression state. According to the neural network calculation method provided by the invention, chromatin accessibility and three-dimensional space interaction data are deeply fused through a dynamic routing module, so that the problem of'black box 'which is inaccurate in prediction and difficult to explain in a traditional deep learning model is solved in a mode of explicitly simulating a real biological regulation mechanism; and a key gene regulatory pathway can be accurately identified.

The invention belongs to the technical field of gene engineering, and particularly relates to application of an OsACX4 gene in regulation and control of rice salt tolerance. The gene related to the rice salt tolerance is identified for the first time and is named as OsACX4, under treatment of different salt concentrations, the salt tolerance of the OsACX4 gene editing function deletion mutant is remarkably improved, and the growth vigor and survival rate of the OsACX4 gene editing function deletion mutant are higher than those of a wild type and an overexpression strain. After salt treatment, the activity of the OsACX4 deletion mutant and ROS removal related enzymes is generally higher than that of a wild type, the ROS cumulant in vivo is lower than that of the wild type, the content of proline for maintaining cell osmotic equilibrium is higher than that of the wild type, and the content of MDA positively related to the cell membrane damage degree is lower than that of the wild type. The results show that the OsACX4 gene negatively regulates and controls the salt tolerance of the rice in the seedling stage, and is a key gene of the rice in response to salt stress in the seedling stage. The method has important application value in the aspects of analyzing a plant salt-tolerant mechanism and breeding salt-tolerant rice varieties.

The invention belongs to the technical field of plant molecular biology, and discloses an application of a soybean GmSW10 gene in reducing soybean plant height, grain weight and grease content and / or increasing protein content. The CDS sequence of the soybean GmSW10 gene is as shown in SEQ ID NO: 1. Knockout of the soybean GmSW10 gene can directly influence the plant height, the grain weight, the oil content and the protein content of soybean seeds, after the soybean GmSW10 gene is knocked out, the weight of soybean seeds and hundred grains is remarkably reduced, the plant height is reduced, the protein content is increased, and the oil content is reduced, which indicates that the GmSW10 gene is a key gene for regulating and controlling the plant height, the grain weight, the oil content and the protein content of the soybean, and the GmSW10 gene can be used for preparing the soybean GmSW10 gene for regulating and controlling the plant height, the grain weight, the oil content and the protein content of the soybean. And a new genetic resource is provided for genetic improvement of soybeans.

The invention discloses a Pseudomonas putida high-yield rhamnolipid fermentation medium based on a step-by-step optimization strategy, an optimization method and application thereof, through the step-by-step strategy of component screening-single-factor optimization-multi-factor interactive optimization, a mixed carbon source and a mixed nitrogen source are taken as cores, and a Plackett-Burman test, a steepest climbing test and Box-Behnken response surface design are combined, so that the high-yield rhamnolipid fermentation medium for the pseudomonas putida high-yield rhamnolipid is obtained. Optimizing to obtain a special fermentation culture medium; two genetic engineering strains estA-2 and estA-3 are constructed by heterologous expression of key genes, and the supply and extracellular transport efficiency of a rhamnolipid synthesis precursor is enhanced; the recombinant strain is applied to the optimized culture medium, and efficient production of a shake flask and a 5L fermentation tank is achieved. The rhamnolipid yield of the estA-3 strain in a 5L fermentation tank reaches 35.59 + / -2.18 g / L, the conversion rate is 0.282 g / g, the productivity is 0.404 g / L / h, the rhamnolipid yield, the conversion rate and the productivity are obviously superior to those in the prior art, and a key technical support is provided for industrial production of rhamnolipid.

The invention discloses a recombinant bacterium for high-yield production of selenomethionine and a preparation method of the recombinant bacterium, and belongs to the technical field of genetic engineering. According to the invention, selenium-rich region (Enshi) soil and withered grass are taken as materials, a bacillus subtilis strain with high selenomethionine synthesis capability is separated and screened, the bacillus subtilis strain is taken as an original strain, and a recombinant strain of high-expression SeMet synthesis key gene thioredoxin reductase (TrxRB) is constructed by utilizing an expression vector pHT43, so that high-yield SeMet engineering bacteria are obtained. The bacillus subtilis rich in selenomethionine and the recombinant bacteria thereof can be used as feed additives, and can be used for supplementing SeMet and improving the feed conversion rate in the production of livestock and poultry and aquatic animals.

The present invention relates to a method for constructing a gene network through a single-cell transcriptome and a method for discovering key genes in differentiation using same, and a composition for the prevention, alleviation, or treatment of colon cancer using the target discovered through the method. The method for constructing a gene network of key genes according to the present invention employs single-cell transcriptome data and thus can be applied to all single-cell transcriptome data. The combination of MYB / HDAC2 / FOXA2 discovered upon application to colon cells can serve as a cancer treatment target that promotes the differentiation of colon cancer cells to revert same into differentiated normal cells.

The invention discloses a method for producing MK-7 through whole-cell regulation and control by using different time sequence type promoters, recombinant escherichia coli and application. According to the method, the time sequence type promoters are used for regulating and controlling expression of different key genes related to MK-7 synthesis. According to the invention, the time sequence promoter is used for whole-cell regulation and control to produce the vitamin MK-7 for the first time, dynamic regulation of the MK-7 is realized, and the production efficiency of the MK-7 can be remarkably improved by accurately regulating and controlling expression of key genes in a synthesis route of the MK-7 and optimizing an intracellular metabolic process. The promoter disclosed by the invention can regulate cell metabolism and enhance synthesis of MK-7 in different growth stages or under different environmental conditions by controlling the time and strength of gene expression. The MK-7 is produced by adopting the constructed recombinant escherichia coli obtained based on whole-cell regulation of the sequential promoter, and the yield of 14 g / L can be finally achieved in a 5L fermentation tank.

The invention belongs to the field of plant genetic engineering, and discloses a key gene OsHPCA1 for positively regulating salt stress and oxidative stress tolerance of rice. The gene (with the login number of Os05g40770) encodes a receptor protein capable of specifically sensing hydrogen peroxide, and belongs to a leucine repeat receptor-like protein kinase (LRR-RLKs) family, wherein the family is the largest subfamily in the receptor-like protein kinase (RLKs). The OsHPCA1 responds to salt and oxidative stress signals, and the salt tolerance and oxidative stress resistance of the rice are positively regulated and responded. The OsHPCA1 overexpression rice strain shows very strong salt tolerance; and the OsHPCA1 knockout strain oshpca1 is more sensitive to salt stress. The OsHPCA1 protein encoded by the OsHPCA1 gene and rich in leucine repeat receptor-like protein kinase can positively regulate and control the activity of catalase C (CatC) in rice and enhance the removal efficiency of H2O2 under salt stress, so that the salt tolerance of rice is improved. When the OsHPCA1 is overexpressed, the activity of CatC in the overexpressed strain OsHPCA1 is obviously increased, the H2O2 removal capability is enhanced, and the salt tolerance and oxidative stress tolerance of the rice are improved.

The invention discloses recombinant escherichia coli capable of producing salidroside and application, exogenous genes are introduced by utilizing a CRISPR / Cas9 technology, the recombinant escherichia coli capable of improving the fermentation yield of the salidroside is obtained by modifying and screening key genes, and finally, the salidroside is obtained by optimizing a fermentation tank process. The recombinant Escherichia coli can reach the highest yield of salidroside produced by fermentation of the scale at present on a 5L fermentation tank. The salidroside is fermented in a 5L fermentation tank for 104 hours, so that the maximum accumulation amount of the salidroside is 33.68 g / L, and industrial enlarged production is facilitated.

The invention discloses a MaSPL1 gene and application thereof in regulation and control of banana fruit after-ripening. The nucleotide sequence of the MaSPL1 gene is as shown in SEQ ID NO. 1. The invention relates to a key gene MaSPL1 for regulating and controlling banana fruit after-ripening in bananas. The invention provides a theoretical basis for genetic improvement of banana fruit after-ripening, has important breeding application value, and can be applied to cultivation of new storage and transportation-resistant banana varieties.

The invention belongs to the technical field of biosynthesis, and particularly relates to a method for regulating and controlling stevioside production through a UDPG synthesis route. According to the invention, a high-efficiency stevioside biosynthesis system is constructed through systematic engineering modification: in the aspect of chassis cell optimization, transcription factors (AtWRKY18, AtWRKY40 and AtMYC2) for regulating and controlling MEP pathway and key genes (dxs and / or dxr) of the MEP pathway are over-expressed, so that supply of terpene precursors is enhanced; and a heterologous MVA pathway (over-expression T7-atoB-mvaS-mvaE-mvK / mvKmut-pmK-mvaD-idi) with a modular design is introduced, so that sufficient supply of the terpenoid precursor is realized. Meanwhile, the intracellular concentration of the UDPG is increased by systematically enhancing a UDPG synthesis pathway (knocking out ugd, glgc, ushA, agp, deoA, tdk, udp, or thrA, and overexpressing galU, pgm, pyrE, pyrH, ugpA, or prs), so that the production capacity of the stevioside is remarkably improved.

The invention discloses a germacene A synthase mutant and application thereof, and belongs to the technical field of enzyme engineering and microbial engineering. Aiming at the bottleneck of the catalytic efficiency of the germacene A synthase, the optimal germacene A synthase mutant DlGASS32H-I97V-Y315F is obtained through semi-rational design based on sequence conservative analysis by taking the germacene A synthase from fungi with a separation wheel layer carbon shell as an object, and the optimal germacene A synthase mutant DlGASS32H-I97V-Y315F is verified through shake flask fermentation in engineering bacteria saccharomyces cerevisiae, so that the yield of beta-elemene is increased by 24%. The combined mutant obtained through a key gene conserved sequence mutation prediction strategy shows remarkably improved catalytic performance, the effectiveness of guiding an enzyme directed evolution strategy based on multi-species sequence conservative analysis is verified, and the yield of germacene A or beta-elemene is remarkably improved. Meanwhile, the method has the advantages of simplicity and convenience in operation, environmental friendliness, mild reaction conditions and the like, and has a very good application prospect in the field of production of germacene A.

The invention discloses application of a gene GlZn2Cys661 in regulation and control of synthesis of ganoderic acid. The nucleotide sequence of the gene GlZn2Cys661 is as shown in SEQ ID NO. 1. The gene expression is regulated and controlled by constructing an overexpression or silent vector, so that the synthesis of the ganoderic acid is promoted or inhibited, and an effective method is provided for regulating and controlling the yield of the ganoderic acid. The invention relates to the technical field of bioengineering, and discloses an application of a gene GlZn2Cys661 in regulation and control of ganoderic acid synthesis. The invention provides a key gene target and a core engineering strain for efficiently producing ganoderic acid through a metabolic engineering means, and has important application value.

The invention discloses a method for high-throughput screening of key genes in a 1, 5-pentamethylene diamine biosynthetic pathway of escherichia coli, and the high-throughput screening is carried out by using a 1, 5-pentamethylene diamine biosensor. The 1, 5-pentamethylene diamine biosensor is a recombinant plasmid obtained by integrating a lysine decarboxylase coding gene cadA and a promoter trc thereof, and a green fluorescent protein (GFP) coding gene and a promoter pyrF thereof into a plasmid skeleton. A fluorescent quantitative flow cytometer is used for screening single-cell thalli with high yield of 1, 5-pentanediamine according to the fluorescence intensity of GFP, key genes can be rapidly screened out from a large number of genes to be screened in a high-throughput mode, and the screening workload and time are greatly saved. According to the application of the key gene in the biosynthetic pathway of the escherichia coli 1, 5-pentanediamine screened by the method disclosed by the invention, the yield of the 1, 5-pentanediamine produced by fermenting the escherichia coli KA30 can be greatly improved, the substrate conversion rate in the fermentation process is improved, the production cost of the 1, 5-pentanediamine is reduced, and the economic benefit is improved.