67 results about "Transporter gene" patented technology

The invention discloses construction and application of an L-malic acid producing issatchenkia orientalis strain, and belongs to the technical field of microorganisms and the technical field of microbial fermentation and metabolic engineering. Through a metabolic engineering means, a cytoplasm metabolic pathway of an issatchenkia orientalis strain is transformed, and a pyruvate decarboxylase coding gene, glycerol dehydrogenase and lactic dehydrogenase are knocked out; a pyruvate carboxylase gene derived from aspergillus flavus, a malate dehydrogenase gene derived from corynebacterium glutamicum and phosphoenolpyruvate carboxylase derived from escherichia coli are screened and integrated, so that a cytoplasm rTCA cycle synthesis pathway is enhanced; inserting a dicarboxylic acid external transporter gene from schizosaccharomyces cerevisiae at a neutral site GD21, and knocking out endogenous dicarboxylic acid internal transporter; the L-malic acid yield of the modified yeast strain in the fermentation process is remarkably increased and reaches 140.16 g / L. The engineering strain provided by the invention has good industrial application potential in efficient L-malic acid production.

The invention discloses an application of a multidrug-resistant efflux transporter gene OsMRET1 in regulation and control of chalkiness traits of rice. A nucleotide sequence of a coding region of the gene is shown as SEQ ID No.1, and a coded amino acid sequence is shown as SEQ ID No.2. The invention further discloses an application of the multidrug-resistant efflux transporter gene OsMRET1 in regulation and control of chalkiness traits of rice. It is found for the first time that the gene OsMRET1 can regulate and control formation of chalkiness, an OsMRET1 gene knockout mutant is created through a gene editing technology, an OsMRET1 gene overexpression strain is obtained through a transgenic technology, it is verified that the OsMRET1 gene is a new gene for regulating and controlling rice chalkiness, and overexpression of the gene can reduce rice chalkiness and improve rice chalkiness. And a new gene resource and a technical route are provided for rice quality improvement.

The invention provides a beta-alanine producing strain as well as a construction method and application thereof. According to the bacterial strain, an acetaldehyde dehydrogenase gene adhE, a lactic dehydrogenase gene ldhA, an acetokinase gene ackA, a pyruvate dehydrogenase gene poxB, a DNA binding transcription inhibition factor coding gene lacI, an aspartate kinase thrA, a pantothenic acid synthase gene panC, an alanine synthesis transaminase gene cycA and an alanine synthesis transaminase gene yfbQ are knocked out from an E.coli W3110 genome, and the bacterial strain is obtained. A phosphoenolpyruvate carboxylase gene ppc, a pyridine nucleotide transhydrogenase coding gene pntAB, an aspartic acid transaminase gene aspC, an aspartic acid decarboxylase gene panD derived from pseudomonas aeruginosa and a beta-alanine transporter gene NCgl0580 derived from corynebacterium glutamicum are integrated at the same time, and the bacillus subtilis is used for producing beta-alanine and has the advantage of high fermentation yield.

The invention discloses a construction method of a cyanobacteria-corynebacterium glutamicum artificial photosynthetic mixed bacteria system. The construction method comprises the following steps: step S1, amplifying by taking plasmid pUC-tho as a template to obtain a theophylline inducible promoter Ptho; step S2, carrying out amplification by taking a Synechocystis sp. PCC 6803 genome as a template so as to obtain a strong promoter Pcpc560, and carrying out amplification on the strong promoter Pcpc560 so as to obtain a strong promoter Pcpc560; step S3, carrying out amplification by taking the plasmid pUC-PAL as a template to obtain an arabidopsis thaliana phenylalanine lyase gene pal; step S4, carrying out amplification by taking the plasmid pUC-cscB as a template to obtain an escherichia coli W sucrose transporter gene cscB; step S5, carrying out amplification by taking the integrated plasmid Pcp3031 as a template, so as to obtain a terminator Trbcl; and step S6, fusing the strong promoter Pcpc560, the arabidopsis phenylalanine lyase gene pal and the terminator Trbcl, and respectively connecting with an expression vector pSI-SPE to obtain the expression vector containing the gene pal. According to the invention, a'cyanobacteria-corynebacterium glutamicum 'artificial photosynthetic mixed bacteria system is constructed by simulating a mutualistic symbiotic relationship ubiquitous in nature, and negative carbon biosynthesis of cis-muconic acid is realized.

The invention discloses a bumblebee candida for efficiently producing sophorolipid as well as a construction method and application of the bumblebee candida. According to the invention, metabolic transformation is carried out on a wild type strain of candida bumblebee, a peroxisome membrane transporter gene PXA1 is knocked out, so that a beta-oxidation pathway of fatty acid competing with a sophorolipid synthesis pathway is moderately inhibited, and on the basis, an endogenous potent promoter pENO is utilized to enhance expression of CYP52M1; therefore, the bumblebee candida engineering bacteria for efficiently producing sophorolipid are constructed. Compared with an original strain, the recombinant candida bumblebee has the advantages that efficient conversion of a fermentation substrate and high yield of sophorolipid can be realized, and the sophorolipid yield of shake flask fermentation reaches 88.50 g / L. The strain is fermented in a 5L fermentation tank and fermented for 7 days after rapeseed oil is added, 232.27 g / L of crude sophorolipid is produced, and high productivity is kept.

The invention discloses a polyamine transporter encoding gene HvPUT5 derived from highland barley and application of the polyamine transporter encoding gene HvPUT5 in improvement of cell osmotic stress tolerance, and relates to the technical field of plant stress resistance molecular breeding and functional gene mining. The HvPUT5 is positioned in cytoplasm, and the over-expression of the gene in a yeast system can obviously improve the survival rate of the yeast under the condition of high osmotic pressure, which indicates that the gene has a positive effect on enhancing the osmotic stress tolerance. The HvPUT5 has a good application prospect, can be used as a candidate gene resource and a molecular marker for drought-resistant breeding, and is applied to genetic improvement and molecular breeding research of crop stress resistance characters.

The invention discloses an application of a soybean GmPT1 coding gene in resisting soybean mosaic virus diseases. The invention discloses application of the soybean inorganic phosphate transporter gene GmPT1 as shown in SEQ ID NO.1 and SEQ ID NO.2 in genetic engineering modification of soybean mosaic virus resistance. The over-expression of the gene can significantly improve the resistance of soybeans to soybean mosaic virus diseases. The soybean inorganic phosphate transporter gene GmPT1 disclosed by the invention can be transformed into soybeans through genetic engineering, and the resistance of the soybeans to soybean mosaic virus diseases is finally regulated and controlled.

Provided are a theanine-producing strain and use thereof in tea fermentation production. A corynebacterium glutamicum is proposed, which includes an alanine decarboxylase CsAlaDC mutant. The theanine-producing strain is obtained by taking the corynebacterium glutamicum as a starting strain, knocking out in sequence an α-ketoglutarate dehydrogenase E1 subunit gene odhA, a glutamate external transporter gene Ncg11221 and a lactate dehydrogenase gene ldh; and / or expressesing a citrate synthase gene gltA, a pyruvate kinase gene pyk and a glutamate dehydrogenase gene gdh; and / or overexpressing an alanine dehydrogenase alaA and integrating a γ-glutamine synthetase GMAS into a cg1960 pseudogene locus of the corynebacterium glutamicum.

The invention belongs to the technical field of microorganism application, and particularly relates to a genetically engineered bacterium for producing L-alanine and application of the genetically engineered bacterium in two-stage fermentation of the L-alanine, and the genetically engineered bacterium takes vibrio natrievibrio as a chassis bacterium and can co-express an alanine dehydrogenase gene and an alanine transporter gene. An alanine dehydrogenase gene is heterologously introduced into a vibrio natriticus genome, and an inactivated gene combination with the optimal L-alanine yield is found out by randomly inactivating a key enzyme in a byproduct production pathway, so that the vibrio natriticus genetically engineered bacterium capable of efficiently producing the L-alanine through a fermentation method is constructed. According to the present invention, the L-alanine is produced through the two-stage fermentation by using the gene engineering bacteria through the temperature-sensitive control strategy so as to reduce the influence of the accumulation of the L-alanine on the cell growth, and the high production of the Xiabanmate is provided. The genetically engineered bacterium is short in growth cycle, glucose can be efficiently utilized to convert the genetically engineered bacterium into L-alanine, the L-alanine is transferred to the outside of cells, separation and purification are simple, cell disruption is not needed, and the yield and conversion rate of the L-alanine can both reach a high level.

The invention relates to a construction method and application of recombinant escherichia coli for increasing the yield of lactoyl-N-tetrasaccharide, and belongs to the field of microbial genetic engineering. According to the invention, three genes tauB, nikD and livM screened by using a transcriptome technology are proved to be capable of promoting the generation of LNT and LNTri II. An engineering strain MG-LNT-09 is used as a starting strain, the biosynthesis efficiency of endogenous transporter genes tauB, nikD, livM, LNT and LNTri II derived from Escherichia coli K-12 derivative bacteria MG1655 is remarkably improved through single plasmid overexpression, the yield of the LNT in shake-flask culture is as high as 7.77 g / L, and the yield of the LNTri II is as high as 3.03 g / L. The optimized strain MG3WB of which the tauB gene is integrated to the hlyE site obtains the maximum LNT titer of 40.35 g / L in a 5L bioreactor without using any antibiotics or plasmids, has a good industrial prospect, and lays a foundation for further industrial production and synthesis of other complex breast milk oligosaccharides.

The present disclosure provides an aggregate of kidney cells having improved expression of transporter genes. Some aspect of the present invention is an aggregate of kidney cells. The aggregate satisfies at least one of the following conditions. (a) The expression level of the OAT1 gene is 1 * 10 <-3 > or more (GAPDH ratio). (b) The expression level of the OAT3 gene is 1 * 10 <-4 > or more (GAPDH ratio). (c) The expression level of the OCT2 gene is 1 * 10 <-3 > or more (GAPDH ratio). (d) The expression level of MATE1 gene is 1 * 10 <-4 > or more (GAPDH ratio). (e) The expression level of the MATE2 gene is 1 * 10 <-5 > or more (GAPDH ratio). (f) The expression level of the MDR1 gene is 1 * 10 <-3 > or more (GAPDH ratio). (g) The expression level of the UART1 gene is 1 * 10 <-6 > or more (GAPDH ratio).

The invention discloses a method for constructing a high-yield natamycin engineering strain through overexpression of an extracellular transport protein gene pimA and a pathway specific regulation gene pimR, and the high-yield engineering strain is applied to fermentation production of natamycin. The yield of the natamycin can reach 19.49 g / L through fed-batch fermentation in a fermentation tank of 5L of the high-yield strain S.ilvosporeus pimA-pimR, and the yield of the natamycin is improved by 33.12% compared with the yield of a chassis strain S.ilvosporeus TUST01; the production intensity is improved by 33.33% compared with the production intensity of a chassis strain S.ilvosporeus TUST01, and the strain has a relatively high industrial application value.

The present invention relates to the field of plant genetic engineering, and specifically to the application of an amino acid transporter gene. This solution reduces the expression level of the OsAAP7 gene in wild-type rice or knocks out the OsAAP7 gene in wild-type rice; the amino acid sequence of the OsAAP7 protein encoded by the OsAAP7 gene is shown in SEQ ID NO.1, or the OsAAP7 protein is a protein with equivalent activity obtained by substituting, replacing, and / or adding several amino acids to the amino acid sequence shown in SEQ ID NO.1. This solution reveals the function of the OsAAP7 gene. By knocking out the expression of the OsAAP7 gene, the elongation of the tiller buds of a single rice plant can be accelerated, the number of tillers per rice plant can be increased, and the number and weight of grains per rice plant can be increased. The OsAAP7 gene can be used in rice breeding to increase rice yield.

This invention provides compositions and treatment methods for reducing hyperphenylalaninemia. [Solution] A genetically modified bacterium is provided, comprising: a) one or more genes encoding phenylalanine ammonia lyase (PAL), which are operably linked to a promoter that is not naturally associated with the PAL gene and can be directly or indirectly induced; b) one or more genes encoding a phenylalanine transporter, which are not naturally associated with the phenylalanine transporter gene and can be operably linked to a promoter that is directly or indirectly induceable; and c) one or more genes encoding mutant fumarate and nitrate reductase (FNR), which are not naturally associated with the FNR gene and can be operably linked to a promoter that is directly or indirectly induceable.

The present invention provides a tea tree-derived transmembrane transporter gene CsMATE33 and an application thereof, belonging to the technical field of biological breeding. The nucleotide sequence of the tea tree-derived transporter gene CsMATE33 is shown in SEQ ID No. 1. Studies have shown that the tea tree-derived transporter gene CsMATE33 provided by the present invention can increase the content of EGCG in yeast, tea trees and tobacco, and provides valuable gene resources for cultivating high-EGCG plants and microbial varieties, and has good application prospects and value.

The invention discloses a genetically engineered bacterium for synthesizing L-glutamine by taking glucose as a carbon source and application of the genetically engineered bacterium. The genetically engineered bacterium takes escherichia coli as a chassis strain; according to the invention, a citrate synthase gene gltA, an aconitic acid hydratase gene can, an isocitrate dehydrogenase gene icd, a glutamate dehydrogenase gene gdh, a glutamine synthetase gene glnA and ammonium transporter genes amt and amtb can be overexpressed. Compared with the prior art, the invention provides a new strain and a new method for biosynthesizing L-glutamine, and the new strain and the new method have the significances of economy, scientificity, low cost, simple process and large-scale production.

The invention discloses recombinant Escherichia coli for producing xylitol by efficiently utilizing L-arabinose without adding glycerol and application, and belongs to the technical field of genetic engineering. Escherichia coli with an araBAD operon and an L-xylulokinase gene lyxK knocked out is used as a starting strain, a glucose specific transporter gene ptsG and a D-xylulokinase gene xylB in a genome are knocked out, and a global regulatory factor CRP is mutated; on the basis of an araA, dpe and lxr three-enzyme expression universal method reported at present, the gene arrangement sequence and copy number in three-enzyme tandem expression are optimized, a promoter and an RBS sequence are optimized and screened, and three-enzyme genes are integrated to a specific position of an escherichia coli chromosome in an optimized multi-copy form. According to the method, the inhibition effect of glucose on L-arabinose is achieved, the in-vivo multienzyme catalysis efficiency of existing synthesis of xylitol from arabinose is remarkably improved, and the yield of precursor arabitol and the yield of the product xylitol are greatly improved.

The invention discloses application of a mulberry calcium transporter MmCAX and a coding gene thereof in manganese stress resistance of plants, the amino acid sequence of the mulberry calcium transporter MmCAX is shown as SEQ ID NO: 2, and the nucleotide sequence of the gene is shown as SEQ ID NO: 1. Through analysis of related physiological and biochemical indexes of mulberry trees under manganese stress and research on the mulberry tree calcium transporter MmCAX, the mulberry tree genome is perfected, and expression analysis and application of the mulberry tree calcium transporter under the manganese stress condition are provided. The mulberry calcium transporter gene MmCAX is transferred into agrobacterium tumefaciens GV3101 by using a VIGS silencing technology, and after gene expression is reduced by using a virus silencing principle, physiological and biochemical indexes of a transgenic plant are detected and the effect of the mulberry calcium transporter gene MmCAX in the aspect of improving the stress resistance of the plant is researched. An important molecular basis is provided for researching a mulberry manganese stress resistance regulation mechanism and cultivating a new variety.

The application discloses an Escherichia coli transmembrane transporter gene and application thereof in vitamin B6 biosynthesis. The application weakens a vitamin B6 transporter gene coding in the Escherichia coli, and the ability of a genetically engineered strain to produce vitamin B6 is greatly improved, and the genetically engineered strain has great application and popularization value. In the best embodiment, the extracellular yield of vitamin B6 of the genetically engineered strain is increased by 33.7% compared with that of a wild type, and the effect is extremely remarkable.

This invention belongs to the field of biological genes and discloses the application of a rice monosaccharide transporter gene OsPLT1 and its transporter in regulating rice growth period. The rice monosaccharide transporter gene... OsPLT1 The nucleotide sequence is shown in SEQ ID NO: 1. This invention also discloses a gene for a rice monosaccharide transporter. OsPLT1 Amplification primers. This invention provides rice. OsPLT1 Novel applications of genes and their encoded proteins in regulating rice growth stage; knockout of wild-type ZH11 using CRISPR / Cas9. OsPLT1 At this time, it can accelerate the growth process of rice without affecting the yield of rice, therefore, OsPLT1 Genes can be used to regulate plant growth periods and increase annual and seasonal yields, and have high economic and application value.

The invention provides a recombinant strain for synthesizing HMB, application of the recombinant strain, and a method for performing whole-cell catalytic synthesis of HMB by using the strain. The recombinant bacterium is obtained by knocking out genes such as a fatty acid metabolism regulation factor fadR gene, a beta-ketoacyl-ACP synthase II gene FabF, a beta-ketoacyl-ACP synthase III gene FabH and the like and enhancing or inserting genes such as a fatty acyl coenzyme A synthetase gene fadD, a long-chain fatty acid transporter gene fadL, a short-chain fatty acid degradation regulation gene cluster gene atoSC and the like. A strain capable of producing HMB by using fatty acid or a substance containing a fatty acid component as a raw material is obtained. The strain has the prospect of green, efficient and industrial production of HMB by utilizing the capability of synthesizing HMB.

The invention relates to a genetically modified rice plant or plant cell, which comprises a heterologous heavy metal ATP enzyme gene operably linked to an OsActin1 promoter, a heterologous ATP binding cassette (ABC) transporter gene operably linked to the OsActin1 promoter, and a heterologous plant chelating peptide synthase gene operably linked to the OsActin1 promoter, wherein the OsActin1 promoter has a lower activity in the seed endosperm of the modified rice plant compared to the activity in other nutritional tissues of the modified rice plant; wherein arsenic (As) and cadmium (Cd) in rice grains of the genetically modified rice plant are reduced as compared to a control rice plant in which the genetic modification is not performed. The invention also relates to a method for constructing such genetically modified rice plants or plant cells, and a kit for carrying out the method.

This invention belongs to the field of crop breeding, specifically relating to an NtLHT1 gene and its application in tobacco's resistance to low nitrogen stress. It involves artificially intervening in the mutation of specific nucleotide sequence fragments in the NtLHT1 gene to obtain two mutated nucleotide sequence fragments, resulting in a mutant NtLHT1 gene, and the application of the mutant NtLHT1 gene in improving tobacco's resistance to low nitrogen stress. The NtLHT1-edited tobacco plants of this invention exhibit enhanced absorption of nitrate and ammonium nitrogen during their vigorous growth period, increased activity of nitrate reductase and glutamine synthase, increased expression of nitrogen transporter genes, and improved nitrogen utilization efficiency. This invention increases the activity of nitrate reductase and glutamine synthase, thereby improving tobacco's resistance to low nitrogen stress and providing a genetic resource and material basis for the development of nitrogen utilization-related genes and strains in tobacco.

The invention belongs to the technical field of insecticides, and particularly relates to a ds-ABC transporter gene for preventing and controlling resistant Frankliniella occidentalis as well as a screening method and application of the ds-ABC transporter gene. The ABC transport protein gene is selected from any one or more of ABCG20, ABCA1, ABCB8, ABCD2 and ABCA5, and the ABC transport protein gene is selected from any one or more of ABCG20, ABCA1, ABCB8, ABCD2 and ABCA5. The screening method comprises the following steps: (1) testing the resistance level of the Frankliniella occidentalis to spinetoram, imidacloprid and thiamethoxam; (2) extracting and purifying the total RNA of the Frankliniella occidentalis population by using a trace total RNA extraction kit, synthesizing cDNA (complementary deoxyribonucleic acid) by taking the RNA as a template, and then carrying out quantitative primer design and gene expression; and (3) carrying out significant analysis on difference analysis of all data through real-time fluorescent quantitative detection, and screening out 5 ds-ABC transporter genes which are subjected to significant up-regulation expression in a resistant population. According to the invention, the key ABC transporter gene for mediating the resistance of the Frankliniella occidentalis to the spinetoram is screened and determined, and a basis is provided for development of a novel anti-resistance insecticide taking the ABC transporter as a target.

The invention discloses a promoter of an alfalfa malic acid transport protein gene MsALMT1 and application of the promoter. The nucleotide sequence of the promoter is shown as SEQ ID No.1. The invention further discloses a preparation method of the promoter. The invention provides the nucleotide sequence of the alfalfa malic acid transporter gene MsALMT1 promoter, and the alkali resistance of the plant can be obviously improved after the promoter drives the overexpression of MsALMT1 in alfalfa, so that a gene resource is provided for the improvement of the alkali resistance of the plant, and the application value is wide.

The invention belongs to the technical field of plant genetic engineering, and particularly relates to a zostera marina magnesium ion transporter gene ZosmaMGT10, a recombinant vector and application. The CDS full length of the gene is 1239 bp, 412 amino acids are encoded, and the gene has a conservative CorA structural domain. It is proved for the first time that ZosmaMGT10 protein has Mg < 2 + > transport activity and can be specifically combined with Mg < 2 + > through E299, E300 and E303 residues, abscisic acid can remarkably induce expression of the ZosmaMGT10 protein in zostera marina leaves, the Mg < 2 + > absorption capacity of zostera marina is enhanced, and then plant stress resistance is improved. The gene ZosmaMGT10 provided by the invention can be used as a key candidate gene, is used for cultivating marine plant varieties with high Mg < 2 + > absorption efficiency and strong stress resistance, and provides a new tool and a theoretical basis for ecological restoration of coastal zones and breeding of stress-tolerant crops.

The application discloses an ABC transporter family gene OsABCA3 and application in plant breeding regulation, belonging to the field of plant genetic engineering. OsABCA3 and application in plant breeding regulation, belonging to the field of plant genetic engineering. OsABCA3 The amino acid sequence of the encoded protein is shown as SEQ ID No: 2. The application realizes the knockout of the ABC transporter gene in rice, OsABCA3 The Cd absorption in the knockout plant under Cd stress is significantly reduced compared with the wild type plant, the Cd content in the grain of the knockout plant planted in Cd contaminated soil is significantly reduced compared with the wild type plant, but the essential element content in the grain and the yield of the rice are not negatively affected, thereby providing a new candidate gene resource for creating safe rice with low Cd content in grain, and providing a potential remediation method for Cd pollution of rice.

The invention belongs to the field of plant genetic engineering, and discloses a trifoliate orange transporter CtrNPF2.1 and application thereof in plant salt resistance and root growth genetic improvement, the CtrNPF2.1 gene is a transporter separated and cloned from citrus rootstock trifoliate orange, and is named as CtrNPF2.1, and the sequence of the CtrNPF2.1 gene is shown as SEQ ID NO.1. The invention further discloses a method for preparing the trifoliate orange transporter CtrNPF2.1 gene. An overexpression vector and an interference vector are respectively constructed by the gene, the gene is introduced into trifoliate orange through agrobacterium tumefaciens-mediated genetic transformation, and biological function verification of an obtained transgenic plant shows that the cloned CtrNPF2.1 gene has the functions of controlling the salt resistance of the plant and regulating the growth of a root system. The development and utilization of the genetic resource are beneficial to reducing the agricultural production cost and realizing environmental friendliness.

The invention discloses an application of a rice phosphorus transporter gene and an encoding protein thereof in improving nutrient utilization efficiency and rice quality. According to the invention, OsPHT1 is mutated or inhibited; 1 gene expression can significantly improve nutrient utilization efficiency and rice taste quality and nutritional quality. The method is mainly embodied in that accumulation of nitrogen and phosphorus nutrients of rice is remarkably reduced, the total protein content, glutelin content, prolamin content, globulin content, albumin content and phytic acid content of rice grains are remarkably reduced, the gel consistency of the rice is increased, the reduction value is reduced, and the eating quality and the nutritional quality of the rice are further improved.

The invention discloses a method for enhancing cholesterol uptake of saccharomyces cerevisiae, and belongs to the technical field of bioengineering. The method comprises the following steps: knocking out transcription inhibition factors MOT3 and ROX1 of genes related to cholesterol uptake and transport of saccharomyces cerevisiae; the transcription activating factors SUT1 and UPC2-1 of the cholesterol transporter gene are over-expressed; expression of sterol esterifying enzyme ARE2 is enhanced, so that uptake and storage of exogenous cholesterol by the saccharomyces cerevisiae are further enhanced. According to the method disclosed by the invention, the uptake and storage of cholesterol by the saccharomyces cerevisiae are enhanced, and an important reference is provided for construction of a steroid hormone synthesis chassis taking cholesterol as a substrate.