31 results about "Synthetic Genes" patented technology

The invention discloses a method for increasing the fermentation yield of mernomycin A based on a reinforced chassis strain glycosyl donor. Specifically, in the heterologous expression process of a sirnomycin A biosynthetic gene cluster in a chassis strain Streptomyces albus J1074, a glycosyl donor synthesis module gene and a resistance gene (MaPBP) are introduced at the same time, and on the basis of improving a glycosyl biosynthetic donor, the physiological tolerance of the chassis strain is also enhanced. Compared with a high-yield strain Streptomyces albus LX03 (CGMCC NO. 20985) of which the tolerance is only optimized, the yield of the strain Streptomyces albus LX02: pJQK610 subjected to genetic engineering optimization is improved by 285%, and the yield reaches 114 mg / L. The strain disclosed by the invention has the advantages that the yield of the strain Streptomyces albus LX02: pJQK610 is improved by 285%, and the yield of the strain Streptomyces albus LX02: pJQK610 is improved by 285%. The invention provides a high-yield technology for improving heterologous expression of the mernomycin A through collaborative optimization of precursor supply and physiological tolerance, and provides reference for efficient production of the medicine.

The present application belongs to the field of biotechnology, and particularly relates to a biosynthesis gene cluster of fungal polyketide compounds and application thereof. The present application provides a heterologous expression method of an aspertaichunol compound biosynthesis gene combination, and a compound aspertaichunol B is obtained through the method, and a compound aspertaichunol A is prepared through acid hydrolysis of the aspertaichunol B, which has important significance for future research and development of an efficient biosynthesis method for preparing the aspertaichunol compound.

The application belongs to the technical field of microbial application, and specifically discloses application of roseofungin Streptomyces roseofunginus in plant disease prevention and treatment. The application finds that after R&M synthesis genes in roseofungin Streptomyces roseofunginus are knocked out, the strain can still synthesize oxazolomycin A. Further finding is that oxazolomycin A shows good inhibitory effect on pepper Phytophthora capsici and Phytophthora sansomei, wherein the EC 50 value of pepper Phytophthora capsici PC35 is 2.95 μg / mL, and the EC 50 value of atractylodes Phytophthora sansomei AMPH-1 is 2.39 μg / mL. The oxazolomycin A prepared by the application can be developed into a biological pesticide that can effectively meet the agricultural development demand of "green ecology", and provides key biocontrol resource support for green prevention and control of plant diseases.

This invention relates to the field of biosynthesis technology, and particularly to engineered yeast strains and their application in the total fermentation production of steviol glycosides. This invention utilizes site-directed mutagenesis to mutate wild-type kaurenoic acid 13-hydroxylase, obtaining the G481L / Q159L mutant. This mutant is then applied to the production of steviol or steviol glycosides, constructing a steviol or steviol glycoside synthesis pathway in the *Saccharomyces cerevisiae* host to achieve efficient production of steviol or steviol glycosides. Simultaneously, by enhancing the level of diterpene precursor synthesis genes in chassis cells (MVA pathway, acetyl-CoA, GGPP, transport proteins, UDPG pathway), the yield of steviol glycosides is further increased, reducing costs for industrial production.

The invention relates to the technical field of bioengineering, in particular to a recombinant halophilic bacterium for producing PHBV by using acetate as a single carbon source and application of the recombinant halophilic bacterium. The expression and / or activity of 2-methyl citrate synthetase and succinate dehydrogenase assembly factor 2 of the recombinant halophilic bacteria provided by the invention are reduced, 3-hydroxyvaleric acid synthetic genes scpA and scpB are overexpressed, an ADP-dependent acetyl coenzyme A synthetase gene acd is overexpressed, and a heterologous monocarboxylic acid transporter gene mctC is expressed. The strain can generate PHBV without adding precursors such as propionic acid and valeric acid, acetic acid or acetate can be used as a sole carbon source to produce PHBV, the yield is close to that when glucose is used as a carbon source, and the problems that the added precursor substances have certain toxicity, so that the growth of thalli is influenced, and the yield is reduced are solved; environmental pollution caused by addition of precursor substances is avoided, and meanwhile, the fermentation production cost of the PHBV is effectively reduced.

The invention provides amycolatopsis mediterranei with gene defects as well as a construction method and application of the amycolatopsis mediterranei. The first aspect of the invention provides a construction method of the amycolatopsis mediterranei with the gene defect, and the construction method comprises the step of knocking out a rifamycin biosynthetic gene cluster in a receptor amycolatopsis mediterranei strain U32 (Cas12a) genome and an integrated plasmid pDZLCas12a containing an FnCas12a coding gene to obtain the amycolatopsis mediterranei with the gene defect. According to the invention, a rifamycin biosynthetic gene cluster in a genome of an amycolatopsis mediterranei strain U32 (Cas12a) and an integrated plasmid pDZLCas12a containing an FnCas12a coding gene are knocked out, so that the amycolatopsis mediterranei with gene defects is constructed, and the amycolatopsis mediterranei can be used as a chassis strain for mining identified or unidentified biosynthetic gene clusters in actinomycetes.

This invention provides Bacillus subtilis ( Bacillus subtle The application of strain A45 in the control of plant diseases, its application in the preparation of microbial agents for the control of plant diseases, and the prepared microbial agents. Bacillus subtilis A45 is deposited at the China General Microbiological Culture Collection Center (CGMCC), with accession number CGMCC No. 33289. The liquid microbial agent containing Bacillus subtilis A45 provided by this invention effectively inhibits the cucumber target spot pathogen, *Clostridium multiflorum* (…). Corynespora cassicola The growth mechanism of this strain lies in its encoding a gene cluster for the synthesis of phytosalicylic acid, purchemin, subtilosin A, and surfactant. Among these, the surfactant synthesis gene cluster is unique to this strain, exhibiting a maximum sequence similarity of 82%. Experiments show that the liquid inoculant containing Bacillus subtilis A45 provided by this invention can effectively inhibit the growth of *Clostridium multiflorum*, the pathogen causing target spot disease in cucumbers. Corynespora cashew nut The growth of ) can be controlled by field application with a prevention and control effect of up to 80%.

The application discloses a rice terpene phytoalexin synthesis gene OsCPS4 and application, relates to the field of plant genetic engineering, and the nucleotide sequence of the gene is shown as SEQ ID NO. 1. The application is a good example that the rice terpene phytoalexin participates in crop insect resistance, has certain reference value for understanding the regulation of the rice terpene phytoalexin on pest resistance, and OsCPS4 the research on the gene provides a good theoretical basis for the molecular mechanism of the rice brown plant hopper resistance gene, and has important significance for molecular design breeding.

The invention belongs to the technical field of plant genetic engineering and the technical field of soybean breeding, and particularly discloses application of a soybean brassinolide inactivating enzyme encoding gene GmBAS1C in regulation and control of the soybean branch number, the CDS sequence of the soybean brassinolide inactivating enzyme encoding gene GmBAS1C is shown as SEQ ID No.1, the amino acid sequence of the encoded protein is shown as SEQ ID No.2, and the amino acid sequence of the soybean brassinolide inactivating enzyme encoding gene GmBAS1C is shown as SEQ ID No.3. The invention further discloses application of the soybean brassinolide inactivating enzyme encoding gene GmBAS1C in regulation and control of the soybean branch number. The gene is located in an endoplasmic reticulum, and formation and development of lateral branches are promoted by up-regulating expression quantities of genes such as brassinolide synthesis genes CPD, ROT3 and BR6OX2, auxin response genes IAA, GH3 and SAUR and the like. The invention also discloses three homozygous gene editing mutants (KOGmBAS1C-1, KOGmBAS1C-2 and KOGmBAS1C-3) of the GmBAS1C gene, a soybean plant containing the gene editing mutants, and a vector construction method of the soybean plant. The invention further discloses the three homozygous gene editing mutants (KOGmBAS1C-1, KOGmBAS1C-2 and KOGmBAS1C-3) of the GmBAS1C gene. Experiments prove that the number of branches of homozygous gene editing mutant phenotypes of the three GmBAS1C genes is remarkably increased. The invention provides a new gene target, a molecular tool and a breeding material for soybean high-branch breeding, and has important agricultural application value.

This invention discloses a genetically engineered bacterium for producing rhamnolipids and its applications, belonging to the field of gene recombination and metabolic engineering technology. The genetically engineered bacterium for producing rhamnolipids disclosed in this invention uses *Yarrowia lipolyticis* ATCC MYA-2613 as the starting strain and heterologously expresses the rhamnolipid synthesis gene. rhlA, rhlB and rhlC, and dTDP-L-rhamnose synthesis gene rmlA, rmlB, rmlC and rmlD This invention discloses a genetically engineered bacterium for producing rhamnolipin, which can efficiently metabolize glucose or glucose-containing cellulose hydrolysates to produce rhamnolipin, achieving a rhamnolipin yield of 42.8 g / L. The genetically engineered bacterium constructed in this invention provides a possibility for the efficient production of rhamnolipin from glucose or glucose-containing cellulose hydrolysates.

The application discloses recombinant saccharomyces cerevisiae for producing wax melia oil and a construction method and application thereof, and belongs to the technical field of biology. By modular integration and optimization of terpene precursor supply and gene expression, the galactokinase genes GAL1, GAL7 and GAL10 are knocked out in the original saccharomyces cerevisiae, and terpene precursor synthesis genes and wax melia oil characteristic terpene synthesis enzyme genes are introduced, so that the recombinant saccharomyces cerevisiae is obtained, that is, a wax melia oil production strain. The proportion of elemol in the fermentation product of the recombinant strain is as high as 51.0%, and six target terpene components are determined. The application provides a stable biological product strain for the application of wax melia oil in the fields of spices, cosmetics and medicines, and has significant economic value and practical application value.

This invention relates to the field of plant genetic engineering technology, and discloses an AtaSQS gene for the biosynthesis of triterpenoids in Aster tataricus and its applications. The AtaSQS gene includes a complete coding region CDS sequence and a promoter sequence, derived from *Aster tataricus* var. *ji*. The CDS sequence is shown in SEQ ID NO.1. The promoter sequence, 1289 bp in length, is located upstream of the CDS sequence and is used to regulate the spatiotemporally specific expression of the AtaSQS gene. This invention cloned the key gene AtaSQS for the biosynthesis of triterpenoids and its promoter sequence from *Aster tataricus* var. *ji*. It clarified its positive regulatory function on the synthesis of total triterpenoids and asterones, and provided related recombinant expression vectors, host bacteria, and breeding methods. This fills the technical gap in the regulatory mechanism of triterpenoid synthesis in Aster tataricus, and can significantly accelerate the breeding process of Aster tataricus varieties with high triterpenoid content, improve their medicinal quality and economic value, and has important application prospects in the fields of plant genetic engineering and the quality improvement of traditional Chinese medicinal materials.

The invention provides a rabbit polyclonal antibody aiming at primary cilia and a preparation method thereof, and belongs to the technical field of biology, and the preparation method comprises the following steps: taking a 200-428 amino acid sequence of human ARL13B protein as a target fragment, synthesizing a gene after codon optimization, cloning the gene to a pet-32a expression vector, and carrying out induced expression in escherichia coli to obtain the rabbit polyclonal antibody aiming at primary cilia. Purifying to obtain high-purity immunogen protein; the nucleotide sequence of the synthetic gene is as shown in SEQ ID NO.1, and the amino acid sequence of the high-purity immunogen protein is as shown in SEQ ID NO.2. Immunizing a New Zealand white rabbit with the high-purity immunogen protein to obtain immune serum; and purifying the immune serum to obtain the rabbit polyclonal antibody. The prepared rabbit polyclonal antibody has higher specificity and signal strength, the titer can reach 1: 64000 or above through ELISA detection, primary cilia can be marked more clearly in a paraffin section immunofluorescence experiment, background noise is remarkably reduced, and the reliability of an experiment result is improved.

This invention relates to the field of gene technology, specifically disclosing a method for enhancing the secretion of secondary metabolites based on the regulation of quorum sensing signal molecules. This method activates the expression of bacterial secondary metabolite synthesis gene clusters through bacterial-fungal interaction, comprising the following steps: (a) co-culturing bacteria and fungi in contact or non-contact mode; (b) during co-culturing, the fungi secrete the quorum sensing signal molecule farnesol; (c) the bacteria sense farnesol through a two-component system, activating the expression of secondary metabolite synthesis genes. This invention effectively overcomes the gene silencing problem caused by metabolic load or lack of regulation in natural strains by activating the originally trace expression of secondary metabolite synthesis gene clusters in bacteria using the fungal-derived quorum sensing signal molecule farnesol.

This invention discloses the chitin synthase 1 (LmCHS1) gene of the leafminer moth, its dsRNA, its synthesis method, and its insect-resistant applications. Through transcriptome analysis, specific primer pairs for the open reading frame of the LmCHS1 gene were designed and synthesized. The LmCHS1 gene sequence was obtained through gene cloning (nucleotide sequence: SEQ ID NO: 1, amino acid sequence: SEQ ID NO: 2). The gene fragment with sequence SEQ ID NO: 3 was selected as the RNAi target sequence. Primer pairs SEQ ID NO: 6 and SEQ ID NO: 7 with restriction enzyme sites were designed and synthesized. PCR amplification was performed, and a prokaryotic expression vector was constructed to induce dsRNA expression. Feeding leafminer moth larvae with dsRNA interfered with LmCHS1 gene expression. After 24 hours, abnormalities were observed in the larvae's body surface, with some larvae failing to complete molting or dying due to an inability to form a normal body surface after molting. After 96 hours, the larval mortality rate was 72.22%, compared to 40% in the GFP control group, demonstrating a significant effect. This invention has broad application prospects in the control of the leaf borer pest in forestry due to its high efficiency, specificity, and safety.

The present invention refers to the heterologous production in microorganisms of modified versions of a predicted isoform of the surfactant protein Lv-ranaspumin-1 (Lv-Rsn-1), whose sequence was inferred from analyzes of the protein extract of the nest foam from the Northeastern Pepper Frog (Leptodactylus vastus). More specifically, it refers to two surfactant proteins that consist of modified versions of the predicted isoform of Lv-Rsn-1; to two synthetic genes each encoding one of these modified versions of the predicted isoform of Lv-Rsn-1; to two expression cassettes each containing one of the synthetic genes encoding one of the modified versions of the predicted isoform of Lv-Rsn-1; to two expression vectors each containing one of the synthetic genes encoding modified versions of the predicted isoform of Lv-Rsn-1; and to two transgenic microorganisms, a bacterium and a yeast, each transformed with one of these synthetic genes and heterologously producing one of the modified versions of the predicted isoform of Lv-Rsn-1. Lv-Rsn-1 has surfactancy, emulsification and dispersancy properties, among others, and its heterologous production allows it to be used in various applications and industrial products, without the need to extract it from the frog nest foam.

This invention belongs to the field of microbial technology, specifically relating to a Brucella-deficient strain 104MΔBioZ, its construction method, and its applications. The Brucella-deficient strain 104MΔBioZ is formed by knocking out a gene involved in biotin synthesis in the Brucella 104M genome. BioZ The strain obtained after gene sequencing; the strain involved in biotin synthesis BioZ The nucleotide sequence of the gene is shown in SEQ ID NO:1. This invention utilizes the knockout of biotin synthesis in Brucella 104M. BioZ A mutant strain was obtained through gene synthesis. The resulting mutant strain exhibited significantly reduced virulence compared to Brucella 104M and demonstrated highly effective resistance to infection by the parent Brucella. Results showed that immunizing healthy mice with the 104MΔBioZ strain described in this invention for brucellosis prevention is safe and provides high protection. The Brucella-deficient strain 104MΔBioZ has broad application prospects in the field of brucellosis prevention and control.

The invention discloses a production method and novel application of tryptophol. Streptomyces longsheng and candida albicans are co-cultured on a solid culture medium to obtain tryptophol. The compound has remarkable anti-tumor activity, IC50 of the compound to human lung cancer cells, mouse colon cancer cells, human liver cancer cells and human breast cancer cells is 154.5 mu g / mL, 144.3 mu g / mL, 122.6 mu g / mL and 110.7 mu g / mL respectively, and meanwhile the compound has important value as a key intermediate of multiple drugs. According to the method, the tryptophol is produced by utilizing a microbial synthesis method, the yield of the tryptophol is increased, the tryptophol is successfully expressed by introducing a tryptophol synthesis gene into escherichia coli C41, and the tryptophol yield is increased to 149.76 mg / L after a tryptophol synthesis path is further optimized. According to the invention, microbial co-culture and genetic engineering technologies are combined for tryptophol synthesis, and the method has profound significance for producing tryptophol through microbial fermentation and promoting the conversion of tryptophol synthesis to a green and energy-saving direction.

The invention relates to a method for cultivating low-antinutritional factor and high-nutrition medicago sativa and application, and belongs to the technical field of plant molecular biology. The method comprises the following steps: firstly, determining key targets for regulating lignin synthesis (COMT1), concanavalin synthesis (ARG10 gene) and plant type (MsCLE3a gene) in medicago sativa by utilizing genomics; then, a multi-target CRISPR-Cas9 gene editing carrier is constructed, and medicago sativa is transformed through an ultrasonic-assisted agrobacterium method; finally, an intelligent phenotypic platform integrating artificial intelligence and hyperspectral imaging is utilized, and excellent mutants with the content of lignin and concanavalin remarkably reduced and compact plant types are screened out in a high-throughput mode. The obtained new variety of medicago sativa is high in safety and high in nutrition density, and stem and leaf powder of the medicago sativa can be widely applied to food and nourishment industries as a super nutrition enhancer.

The invention discloses an OMV vaccine capable of preventing group B streptococcus III serotype infection and a preparation method thereof. The invention belongs to the field of synthetic biology, and particularly relates to an OMV vaccine capable of preventing group B streptococcus III serotype infection and a preparation method of the OMV vaccine. The recombinant escherichia coli contains a lipoid A deacylase coding gene, a lipoid A dephosphatase coding gene and a group B streptococcus capsular polysaccharide length control gene coding gene; the gene does not contain an acetyl glucosamine phosphotransferase coding gene, an escherichia coli O antigen synthesis gene cluster coding gene, a lipoid A myristoyl transferase coding gene, an escherichia coli O antigen chain length control gene coding gene, a lipoid A palmitoyl transferase coding gene and an escherichia coli sialic acid degradation gene cluster coding gene. The composition also contains a group B streptococcus III capsular polysaccharide synthesis cluster coding gene. The recombinant escherichia coli can be used for producing an OMV vaccine for preventing group B streptococcus III serotype infection.

The invention provides a tandem gene for biosynthesizing astaxanthin in rape seeds, an expression cassette and an expression method, and belongs to the technical field of gene engineering. The invention provides a tandem gene for biosynthesis of astaxanthin, and four carotenoid anabolism genes are firstly constructed into the tandem gene for a biosynthesis path of astaxanthin. The four carotenoid anabolism genes are driven based on the rape seed specific NAPIN promoter to construct an astaxanthin biosynthesis path in the rape seeds and synthesize the astaxanthin, so that the biosynthesis efficiency of the astaxanthin is improved; the expression of the astaxanthin biosynthetic gene and the accumulation of the astaxanthin in the seeds do not influence the growth, development and reproduction of the rape; in addition, fat-soluble carotenoid and astaxanthin are naturally integrated with grease in the rape seeds, and astaxanthin rapeseed oil is directly used as a product, so that the activity of astaxanthin is maintained, and the later astaxanthin extraction production cost can be reduced.

The invention relates to direct cloning of a biosynthetic gene cluster, in particular to a direct cloning method and application of an ultra-large biosynthetic gene cluster, a microbial genome is extracted from a low-melting-point agarose gel block through combination, and the ultra-large gene cluster can be rapidly cloned in a targeted manner by utilizing the specific cutting characteristic of CRISPR / Cas and a saccharomyces cerevisiae TAR technology. According to the method, mechanical shearing can be avoided by extracting and treating the genome in the thallus embedding block, and the operation is simple, so that the integrity of an ultra-large target gene cluster can be maintained to the maximum extent, and particularly, the probability that rearrangement is extremely easy to occur in a yeast body due to excessive overlapped fragments can be reduced for (ultra)-large PKS / NRPS gene clusters. CRISPR / Cas mediated cleavage can occur at any DNA site, and a target gene cluster can be released in a most accurate and fastest manner, so that targeted cloning is not limited by restriction enzyme cleavage sites any more. The high efficiency of the TAR technology enables the target cloning of (ultra) large gene clusters to be simple and feasible.

This invention relates to the field of air disinfection devices, specifically to a composite intelligent circulating air disinfection device and method. The device includes a housing, and within the housing are a gas control device, a liquid control device, a light control device, a photoexcitation device, and a nanoenzyme adsorption device. This invention integrates modules corresponding to four methods: electrochemical strong oxidizing bactericidal liquid disinfection, ultraviolet irradiation disinfection, nanoenzyme disinfection, and photoexcitation disinfection. This enables highly efficient killing of various pathogens in the air, including bacteria, fungi, and viruses. Furthermore, through ultraviolet-assisted electrochemical oxidation technology, it downregulates the expression of bacterial cell wall synthesis genes and efflux pump genes, inhibits energy metabolism, and induces key gene mutations, effectively degrading and inactivating antibiotic resistance genes, blocking the spread of drug resistance. This significantly improves the quality and efficiency of air disinfection while ensuring the coexistence of humans and the machine.

The present invention concerns bioluminescent plants and methods for producing them. Among said methods, the invention relates, in particular, to a method of introducing luciferase genes and luciferin biosynthesis genes into the chloroplasts of a plant. In addition, the invention concerns a method of producing light comprising adding chemical compounds to a culture medium of the plant according to the invention.

The present invention relates to a method for producing recombinant proteins in which cells have a more stable heterologous protein expression environment by knocking down or knocking out intracellular proteolytic enzyme genes (including but not limited to ATP-dependent serine protease lon, serine endopeptidase htrA or a combination thereof), the intracellular synthesis of difficult-to-express or low-expression recombinant protein which is easily degraded by a host is facilitated. Furthermore, by regulating and controlling morphological related genes (including but not limited to a microbial skeleton protein gene mreBCD, a cell division ring synthesis gene ftsABEIQWXYZ, a cell division suppression protein gene sulA and a cell division ring positioning gene minCDE), larger cell morphology and volume are generated, so that more intracellular proteins can be accommodated; therefore, the production of recombinant proteins (such as lipase, lactoferrin LTF, alpha-lactalbumin alpha-LA and sweet proteins (such as Monellin) and single-cell proteins is improved.

The application belongs to the technical field of plant genetic engineering, and particularly relates to application of GhDWARF27 gene in regulation of plant verticillium wilt resistance and breeding of verticillium wilt resistant plants. The application discloses that application of GR24, an artificial synthetic analogue of strigolactone (SLs), or overexpression of a SLs biosynthesis gene GhDWARF27 can improve the resistance of plants to verticillium wilt, and silencing of the GhDWARF27 gene can result in reduction of SLs content in roots of the plants and reduction of the resistance of the plants to verticillium wilt. SLs can activate expression of JA response related genes through JA and ABA biosynthesis and signal pathways, and can enhance the resistance of the plants to verticillium wilt in an ABA dependent manner by enhancing the activity of antioxidant enzymes. The technical scheme and molecular mechanism of SLs for enhancing the resistance of cotton to verticillium wilt will provide a new approach for cotton breeding and prevention of cotton verticillium wilt.

This invention discloses a strain of Bifidobacterium animalis with both lipid-lowering and anti-aging effects, and its application in enhancing the efficacy of licorice through transformation, belonging to the fields of microbial technology and pharmaceutical technology. This strain can ferment licorice extract, increasing the content of active ingredients such as glycyrrhetinic acid, glycyrrhetinic acid 3-O-glucuronide, and 5-O-methyl genistein, thereby enhancing its efficacy. Specifically, it inhibits weight gain and reduces four blood lipid levels under a high-fat diet; reduces the levels of aging-related secretory phenotypes IL-1β and IL-6; reduces the content of hepatic β-galactosidase; reduces damage caused by hepatic lipid accumulation; and promotes the expression of lipid metabolism genes MGL, HSL, ATGL, and PPAR-α, while inhibiting the expression of lipid synthesis genes PPAR-γ and Fasn. Therefore, the combined application of Bifidobacterium animalis subsp. lactis CCFM1363 and licorice extract has broad potential for the development of lipid-lowering and anti-aging products.

This invention belongs to the field of edible fungi genetic breeding technology, and relates to a Cordyceps militaris strain that does not produce aspergillin, with the preservation number CGMCCNo.40266. This strain does not contain the CCM_02059 gene, CCM_02060 gene, or any of the CCM_02059 to CCM_02060 genes. It also relates to a method for creating this strain through CRISPR / Cas9 gene editing, comprising the following steps: S1, determining and synthesizing the sgRNA nucleotide sequence of the target gene; S2, determining and synthesizing the nucleotide sequences of the upstream and downstream homologous arms of the target gene; S3, ligating the sgRNA nucleotide sequence described in S1 and the homologous arm nucleotide sequences described in S2 into the pAMA1-Cas9-hyg vector to construct a knockout vector; and S4, transforming the knockout vector described in S3 into Cordyceps militaris protoplasts via PEG-mediated transformation to obtain a Cordyceps militaris strain with the target gene knocked out. This invention utilizes CRISPR / Cas9 editing technology to edit the gene cluster of inoculin synthesis in Cordyceps militaris, disrupting the inoculin synthesis pathway and breeding Cordyceps militaris strains that do not produce inoculin and have no foreign gene insertions, which is of great significance to the development of the Cordyceps militaris industry.

The application provides a kind of recombinant corynebacterium glutamicum for producing beta- arbutin and its construction method and application, belong to genetic engineering technical field, the recombinant corynebacterium glutamicum for producing beta- arbutin, is inactivated polyketide synthase gene on the basis of corynebacterium glutamicum again expression beta- arbutin synthesis gene after being obtained.The application inactivates polyketide synthase gene in corynebacterium glutamicum, and successfully obtains the strain 13032- Δpks13 with enhanced biofilm formation ability, can effectively enhance the adsorption growth ability of corynebacterium glutamicum cell on the surface of various media, overexpresses beta- arbutin synthesis gene in 13032- Δpks13 strain as fermentation strain, to realize the application of high-efficiency production of beta- arbutin in adsorption immobilization mode.

The invention provides engineering vibrio natriticus for producing farnesene by using ethanol and a preparation method of farnesene, and relates to the technical field of bioengineering. In the embodiment of the invention, traditional chassis organisms such as yeast which grow slowly are abandoned, and the vibrio natriticus of which the doubling time is less than 10 minutes is used as a chassis; and engineering vibrio natrieti capable of efficiently utilizing ethanol is obtained by combining directed evolution, so that the thallus proliferation and fermentation period is greatly shortened. The selected exogenous plasmids Kesling 2187 and pFZ71 carry a gene cluster with a definite function, and after the gene cluster is successfully introduced into the vibrio natriticus, engineering vibrio natriticus with stable inheritance is formed. MVA pathway key genes (atoB, tHMG1 and the like) carried by the exogenous plasmids and a farnesene terminal synthesis gene (AFS1) have a synergistic effect, so that a complete and efficient synthesis pathway from ethanol to acetyl coenzyme A to FPP to alpha-farnesene is formed, the metabolic flux loss is reduced, and the farnesene synthesis flux is improved.