2094 results about "Engineered genetic" patented technology

The invention relates to the technical field of genetic engineering, and particularly discloses a genetic engineering bacterium for producing melanin as well as a construction method and application of the genetic engineering bacterium. According to the application, the yield of the melanin can be effectively increased by modifying a metabolic pathway of the genetically engineered bacterium, and the yield of the melanin reaches 6.2 g / L and 18.5 g / L respectively through shake-flask culture and 5L fermentation tank culture. Compared with the prior art, the yield of melanin produced by the genetically engineered bacterium is improved by 30% at the shake flask fermentation level, the fermentation yield of a fermentation tank is improved by 4.3 times, in addition, expensive tyrosine is changed into glucose as a fermentation precursor substance, the production cost is greatly reduced, and an efficient and feasible solution is provided for large-scale production of melanin.

Provided are S-adenosylmethionine synthetase mutants and the use thereof, belonging to the technical field of biological enzyme engineering. Site-directed combinatorial mutations are performed on positions 189, 200, 234, 266 and 371 of S-adenosylmethionine synthetase MAT to obtain a series of mutants with improved enzyme activity, wherein the specific enzyme activity of I189V / V266H and V266H / N371R is increased by more than 10 times relative to the parent. The yields of S-adenosylmethionine synthesized by whole cells of recombinant bacteria expressing mutants I189V / V266H / N371R, V266H / Q234N / N371R and I189V / V266H / Q234N / N371R all reach 1550 mg / L or more. The provided enzyme mutants have broad application prospects in the production of S-adenosylmethionine and the construction of genetically engineered microorganisms for S-adenosylmethionine.

The invention discloses a recombinant XVII type collagen engineering bacterium as well as a preparation method and application thereof, and belongs to the technical field of genetic engineering, and the recombinant XVII type collagen engineering bacterium is realized through three steps of designing and optimizing collagen, improving translation through codon optimization and selecting a vector for cloning. By optimizing the amino acid sequence and the codon, the expression efficiency and the stability of the recombinant XVII type collagen in engineering bacteria are improved. The optimization not only improves the production efficiency of the protein, but also reduces the production cost. The recombinant XVII type collagen has potential in the medical field, especially in the fields of skin repair, wound healing, aging resistance and the like. Meanwhile, in the fields of beauty and biological materials, collagen is widely applied to products such as masks and skin care products as a basic component, and can promote skin cell regeneration and enhance skin elasticity and repair capacity.

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

The invention belongs to the technical field of plant genetic engineering, and particularly relates to an SCRE10 gene for improving rice disease resistance and application of the SCRE10 gene, the base sequence of the SCRE10 gene is shown as SEQ ID NO.1, and the amino acid sequence of the SCRE10 gene is shown as SEQ ID NO.2. Through construction of a dexamethasone induced expression SCRE10 transgenic rice plant, it is found that the transgenic rice can significantly induce PR gene expression and active oxygen outbreak, and the resistance of rice to false smut and bacterial leaf blight can be improved. The invention proves that heterologous inducible expression of the Ustilaginoidea virens SCRE10 gene has the function of positively regulating the disease resistance of the rice, and the SCRE10 gene can be used for improving the disease resistance of the rice, which is of great significance to the creation of disease-resistant germplasm of the rice.

The invention belongs to the technical field of microbial genetic engineering, and particularly relates to an alpha-2, 3-sialyltransferase mutant and application thereof. The a-2, 3-sialyltransferase mutants V175N, P198A, G173D, D174Q, F196K and A315H are obtained by performing site-specific mutagenesis on six amino acids in an a-2, 3-sialyltransferase amino acid sequence coded by a gene derived from Campylobacter jejuni (OH4384), the yield of 3 '-sialyllactose is remarkably increased by single-point mutagenesis of the V175N, the yield of 3'-sialyllactose is remarkably increased by single-point mutagenesis of the V175N, the yield of 3 '-sialyllactose is remarkably increased by single-point mutagenesis of the P198A, the G173D, the D174Q, the F196K and the A315H, the optimal recombinant microbial strain 3 '-SL-09 (V175N / P198A / D174Q) is obtained through combination of multiple mutation sites, the yield can reach 5.91 g / L, the bottleneck problem of low catalytic efficiency of sialyltransferase is effectively solved, and an enzyme element with excellent performance is provided for high-efficiency and low-cost biological manufacturing of 3'-SL.

The invention provides a genetically engineered bacterium for producing O-succinyl-L-homoserine as well as a construction method and application of the genetically engineered bacterium. In a chassis bacterium genome, the expression of a 2-ketoglutaric acid decarboxylase encoding gene sucA is enhanced, and the expression of a succinyl-coenzyme A synthetase encoding gene sucD is weakened, so that the supply of succinyl-coenzyme A is increased; the method comprises the following steps: increasing the NADPH (Nicotinamide Adenine Dinucleotide Phosphate) reducing capacity and ATP (Adenosine Triphosphate) energy supply of a chassis bacterium, increasing DNA (Deoxyribose Nucleic Acid) in combination with a transcription dual regulatory factor ompR to improve the stress resistance of escherichia coli under high osmotic pressure, and introducing an overexpression plasmid containing a homoserine transsuccinylase coding gene metA to construct the genetically engineered bacterium for producing O-succinyl-L-homoserine. The engineering strain obtained through a systematic metabolic engineering modification strategy can realize effective accumulation of OSH, the shake flask yield of OSH reaches 19.8 g / L, the fed-batch fermentation yield of a 5L fermentation tank reaches 110.5 g / L, the sugar-acid conversion rate reaches 52.6%, and a foundation is laid for subsequent construction of high-yield OSH engineering bacteria.

The invention relates to the field of rice gene engineering, and discloses an application of a GSE3.1 protein and a coding gene thereof in regulating and controlling the grain length, the grain width and the grain weight of rice seeds. According to the invention, a GSE3.1 gene is edited based on a CRISPR / Cas9 technology, a japonica rice variety Zhonghua 11 (ZH11) is introduced by using an agrobacterium-mediated method, and a knockout mutant is obtained through screening. The homozygous knockout mutation of the GSE3.1 gene in the invention leads to reduction of the grain width and grain length of rice seeds and reduction of thousand grain weight. According to the invention, the constructed plant overexpression vector ProActin: GSE3.1 is expressed in wild type ZH11, so that compared with the wild type plant, the seed grain length and grain width of the transgenic plant are obviously increased, and the thousand grain weight is obviously increased. Therefore, the GSE3.1 gene and the encoding protein thereof can regulate and control the size and the weight of the rice seeds, and are of great significance to cultivation of high-yield rice varieties.

The invention discloses a spermidine synthetase mutant and application of the spermidine synthetase mutant in synthesis of spermidine, and belongs to the technical field of genetic engineering and metabolic engineering. According to the method, the spermidine synthetase is mutated firstly, the efficiency of the spermidine synthetase is improved, and on the basis, the dissolved oxygen and fed-batch concentration are optimally controlled through fermentation, so that the yield of spermidine is improved. The corynebacterium glutamicum provided by the invention is an engineering bacterium with plasmids and without defects, and after the corynebacterium glutamicum is subjected to fed-batch fermentation in a 5L fermentation tank for 68 hours, the yield of spermidine reaches 25.1 g / L, and the sugar-acid conversion rate is 17.9%.

The invention discloses application of Indel marker dosage typing in detection of sweet potato soft rot resistance, and belongs to the technical field of plant genetic engineering. The Indel molecular marker related to the sweet potato soft rot resistance is obtained through screening, the nucleotide sequence of the Indel molecular marker is shown as SEQ ID NO.2, and an 8bp sequence is inserted into or deleted from the 161st site of the sequence shown as SEQ ID NO.2. Experiments find that target bands of amplification products are obtained through amplification of the Indel molecular marker and electrophoresis, dosage values of two genotypes are obtained according to gray values of the dyed electrophoretic bands, and results show that the insertion type Indel molecular marker is positively correlated with the sweet potato soft rot resistance. The method is of great significance to the dose effect research and molecular assisted breeding of sweet potato gene functions.

The invention relates to the field of rice gene engineering, and discloses an application of OsMYB36 protein and a coding gene thereof in regulation and control of rice salt tolerance. The gene of the OsMYB36 protein is edited on the basis of a CRISPR / Cas9 technology, an agrobacterium-mediated method is utilized to introduce a japonica rice variety Zhonghua 11, and a knockout mutant is obtained through screening; tests prove that the survival rate of the rice mutant with the OsMYB36 gene knocked out is remarkably higher than that of a wild-type Zhonghua 11 plant after the rice mutant is subjected to stress treatment for 11 days by 150 mM NaCl and 200 mM NaCl salt and then rehydrated for 7 days after the rice mutant is subjected to two-leaf and one-core stage. And a simple and effective technical means is provided for rapidly creating a new salt-tolerant rice strain. Therefore, the OsMYB36 protein and the coding gene thereof disclosed by the invention can regulate and control the salt tolerance of the rice, and are of great significance to the cultivation of salt-tolerant transgenic rice.

The invention relates to the technical field of synthetic biology, metabolic engineering and cell culture. The present invention provides a cell for the production of a negatively charged, preferably sialylated, oligosaccharide wherein the cell is genetically engineered to have or express, preferably overexpress, a hydrolytic UDP-N-acetyl-D-glucosamine-2-epimerase. The invention further provides the use of said cells in culture or incubation. Also described are methods of producing negatively charged, preferably sialylated oligosaccharides using the cells, and purification of the negatively charged, preferably sialylated oligosaccharides.

The invention discloses a genetically engineered bacterium and application of the genetically engineered bacterium in preparation of ergothioneine. The genetically engineered bacterium carries genes egtB, egtD and egtE for coding ergothioneine synthetase, and the genes egtB, egtD and egtE are assembled with a protein scaffold assembly SGP; the protein scaffold assembly SGP comprises a protein scaffold GBD, a protein scaffold SH3 and a protein scaffold PDZ. The inventor finds that the ergothioneine synthetases egtB, egtD and egtE are assembled by using the protein scaffold assembly SGP, so that the ergothioneine can be efficiently synthesized, and the protein scaffold assembly has an important application prospect.

The invention belongs to the technical field of genetic engineering and enzyme engineering, and particularly relates to a genetically engineered bacterium for stably and efficiently producing L-isoleucine as well as a construction method and application of the genetically engineered bacterium. The genetically engineered bacterium takes E. coliTHRD as a host and contains an RNA (Ribonucleic Acid) polymerase beta-subunit mutant coding gene rpoBM; compared with a parent RNA polymerase beta-subunit, the RNA polymerase beta-subunit mutant has the advantage that the amino acid residue at the 618th site of the amino acid sequence is sequentially replaced with leucine from the N terminal to the C terminal. The genetically engineered bacterium disclosed by the invention can tolerate L-isoleucine and accumulate less alpha-ketobutyric acid, and has a very good industrial application prospect.

The invention discloses a glutathione bifunctional synthetase mutant with improved catalytic activity and application, and belongs to the field of gene engineering and fermentation engineering. A series of mutants with improved enzyme activity are obtained by carrying out site-specific combinatorial mutation on a site 61, a site 136, a site 485, a site 498 and a site 722 of the glutathione bifunctional synthetase GshFst, the specific enzyme activity reaches 13.58 U.mg <-1 >-27.34 U.mg <-1 >, the half-life period at 37 DEG C reaches 195-320 min, and the enzyme activity and the thermal stability are remarkably improved compared with those of a wild type. The enzyme mutant provided by the invention has a wide application prospect in the production of glutathione and the construction of genetically engineered microorganisms for producing glutathione.

The invention belongs to the technical field of bioengineering, and relates to a transaminase mutant, a recombinant genetically engineered bacterium and application of the transaminase mutant in catalytic synthesis of (R)-1-Boc-3-aminopiperidine.The transaminase mutant is obtained by conducting single-point or combined mutation on the 131 site and / or the 197 site of an amino acid sequence shown in SEQ ID NO.2; the mutation sites comprise that the 131 phenylalanine is mutated into aspartic acid, threonine or tyrosine, and / or the 197 lysine is mutated into arginine or leucine. Experimental results show that compared with wild type transaminase, the catalytic activity, the thermal stability and the organic solvent tolerance of the obtained mutants, especially single-point mutants MyTA1-F131Y and MyTA1-K197R and a combined mutant MyTA1-F131Y-K197R, are all remarkably improved, and compared with the wild type transaminase, the catalytic activity, the thermal stability and the organic solvent tolerance of the obtained mutants are all remarkably improved. The mutant MyTA1-F131Y-K197R can be used for efficiently catalyzing asymmetric amination of N-Boc-3-piperidone to synthesize (R)-1-Boc-3-aminopiperidine, the conversion rate of the (R)-1-Boc-3-aminopiperidine after the (R)-1-Boc-3-aminopiperidine reacts for 24 hours under the condition that the substrate concentration is 100 g / L can reach 90% or above, and the mutant MyTA1-F131Y-K197R has a good industrial application prospect.

The invention discloses a Parkinson's disease animal model, a construction method and application, and relates to the field of animal genetic engineering and disease model construction. According to the model, seven mutation sites related to the human familial Parkinson's disease are introduced behind an initiation codon of a fourth exon of an endogenous SNCA gene, and the mutation sites are A18T, A29S, A30P, E46K, H50Q, G51D and A53T and have genotypes related to the human familial Parkinson's disease. The pig model shows nigra dopaminergic neuron reduction and cortical neuron reduction in the newborn period, and the core pathological process of the human Parkinson's disease can be systematically reproduced. The model can be widely applied to Parkinson's disease pathogenesis research, drug screening and cell therapy effect evaluation, and has important scientific research and preclinical application values.

The invention relates to the technical field of plant genetic engineering and genetic breeding, in particular to a method for regulating and controlling thermal morphogenesis of arabidopsis thaliana and application. The method is realized by regulating and controlling an ATG8 gene in arabidopsis thaliana, and the ATG8 gene is a combination of ATG8b and ATG8e, or a combination of ATG8f and ATG8h. The application comprises a thermal morphogenesis response weakening plant obtained through cultivation, or a seed thereof, a propagation material, and a kit for identifying the thermal morphogenesis ability of arabidopsis thaliana and weakening the thermal morphogenesis ability of arabidopsis thaliana. The invention discloses a new function of the ATG8 gene for the first time, and rigorous experiments prove that a plurality of members (ATG8b, ATG8e, ATG8f and ATG8h) of the ATG8 gene family positively regulate thermal morphological establishment of arabidopsis thaliana, so that people's understanding of the effect of autophagy in plant temperature sensing and adaptive growth is expanded. The function of the autophagy core gene ATG8 is combined with the breeding target of crop heat-resistant plant type improvement for the first time, and a brand new technical path is provided.

The invention belongs to the technical field of genetic engineering breeding, and particularly relates to application and a method of a transcription factor CaBTF3 for regulating capsorubin synthesis. According to the capsicum transcription factor CaBTF3 and the coding gene thereof provided by the invention, the biosynthesis of capsorubin can be influenced by adjusting the expression level of a key enzyme gene in a capsorubin synthesis route and regulating and controlling the development and quantity of fruit chloroplast, the expression of the CaBTF3 gene in capsicum fruits is inhibited, the accumulation amount of capsorubin in the capsicum fruits can be reduced, and the yield of capsorubin is increased. The color of the pepper fruit is further influenced. By enhancing the expression level of the CaBTF3 gene, the content of capsorubin in capsicum fruits can be increased, the gene is applied to plant genetic engineering breeding, a new effective way is provided for creating capsicum varieties with high capsorubin content, and the gene plays an important role in improving the economic benefits of the capsicum industry and popularizing and applying the capsorubin.

The invention belongs to the technical field of biology, particularly relates to the technical field of plant genetic engineering, and more particularly relates to application of an MYC gene in regulation and control of chloroplast development and composite stress resistance. Specifically, the invention innovatively discovers that the combined action of warm and JA can significantly induce seedling cotyledon chloroplast dysplasia, and deletion / reduction of MYC gene expression can significantly inhibit chloroplast dysplasia of plants under the combined action of warm and JA. The discovery provides guidance for remarkably improving plant seedling chloroplast development in a complex dynamic environment and improving the seedling survival rate and photosynthetic productivity, can promote optimization of a chloroplast artificial regulation and control technology, and realizes directional design of efficient photosynthetic organs in facility agriculture and an artificial photosynthetic system.

The invention belongs to the technical field of gene engineering, and particularly relates to an acid protease mutant as well as a coding gene, a preparation method and application thereof. The amino acid sequence of the acid protease mutant is as shown in SEQ ID NO. 2. The acid protease mutant provided by the invention has the advantages of high enzyme activity, strong pH and temperature adaptability, good high temperature resistance and the like, can react in a wide pH value (pH 2.0-6.0) and temperature range (30-70 DEG C), is beneficial for improving the flexibility of a production process, particularly can adapt to industrial production at a relatively high temperature and can meet a relatively long-time high-temperature catalytic environment, and the acid protease mutant has a wide application prospect. The enzyme prepared by the method is high in enzyme activity, not easy to inactivate in a high-temperature environment, can maintain 70% or more of enzyme activity for 1 h at 50 DEG C, can maintain 55% or more of enzyme activity for 1 h at 55 DEG C, and shows good market prospects and industrial application values in the fields of wine brewing, feed processing, leather softening and the like.

The invention discloses a mint McTTG1 gene as well as an expression protein and application thereof, and belongs to the technical field of plant genetic engineering. The nucleotide sequence of the mint McTTG1 gene disclosed by the invention is shown as SEQ ID NO.1, the amino acid sequence of the expression protein of the mint McTTG1 gene is shown as SEQ ID NO.2, and the mint McTTG1 gene belongs to WD40 family genes participating in regulation and control of epidermal hair development and synthesis of anthocyanin and procyanidine. Transgenic experiments show that after the McTTG1 gene is over-expressed in the arabidopsis thaliana mutant ttg1-13 with epidermal hair loss and significantly reduced accumulation of anthocyanin and procyanidine, the formation of the epidermal hair on the surface of the mutant can be recovered, and the accumulation of anthocyanin and procyanidine can be promoted. Gene resources are provided for plant variety improvement, and cultivation of new mint varieties is facilitated.

The invention belongs to the technical field of plant genetic engineering, and particularly relates to application of a biological product for overexpressing an OsARF7 gene to improvement of rice disease resistance, the nucleotide sequence of the OsARF7 gene is shown as SEQ ID NO.1, and the amino acid sequence of the OsARF7 gene is shown as SEQ ID NO.2. According to the invention, an OsARF7 overexpression rice strain is constructed by means of transgenosis. Tests show that the overexpression of the OsARF7 can significantly enhance the immunocompetence of the rice, for example, the expression level of a disease-resistant related gene is improved, active oxygen outbreak is significantly induced, and the resistance of the rice to false smut and bacterial leaf blight is significantly improved. The invention proves that the OsARF7 has a positive regulation effect on rice immunity, can be used for improving the disease resistance of rice, and has important significance for creating disease-resistant rice germplasm resources.

This invention provides a genetically engineered bacterial strain, its preparation method, and its application in the synthesis of perillaldehyde. The genetically engineered bacterial strain includes the CcLPPS gene derived from *Cistus creticus*; the strain is selected from *Yarrowia lipolytica*. This invention constructs a stable, highly efficient de novo perillaldehyde synthesis strain. The entire fermentation process requires no addition of any organic reagents or intermediate substances, significantly reducing production costs and simplifying subsequent separation and purification steps. The engineered *Yarrowia lipolytica* strain provided by this invention exhibits high production intensity and a simple fermentation process, showing promising application prospects and laying the foundation for the microbial fermentation production of perillaldehyde.

The invention relates to the technical field of biological genetic engineering, in particular to application of a brassica napus BnaA01G0331000ZS gene in drought stress resistance. The BnaA01G0331000ZS gene is cloned by using cDNA of double 11 leaves as a template through a sequence published in a brassica napus genome database, an over-expression vector is constructed and genetically transformed into rape variety high oil for transgenic function verification, the drought resistance of the obtained over-expression transgenic line is obviously enhanced compared with that of a wild type, and the BnaA01G0331000ZS gene has the advantages that the BnaA01G0331000ZS gene is obtained; therefore, the gene can positively regulate the drought stress resistance of the rape. Therefore, the invention provides a new gene resource for adversity stress resistance of rape.

Genetically engineered immune cells, which express at least two metabolism modulating polypeptides and optionally a chimeric receptor polypeptide (e.g., an antibody-coupled T cell receptor (ACTR) polypeptide or a chimeric antigen receptor (CAR) polypeptide) capable of binding to a target antigen of interest. Also disclosed herein are uses of the engineered immune cells for inhibiting cells expressing a target antigen in a subject in need thereof.

The invention belongs to the technical field of enzyme catalysis, and particularly relates to a dehydrogenase mutant, a gene, an engineering bacterium and application, the dehydrogenase mutant is obtained through mutation of original dehydrogenase, the sequence of the original dehydrogenase is SEQ ID NO.1, and the mutation mode is that the 344 site is mutated from A to W, K, F, S or N, or the 130 site is mutated from H to T, W or F; the dehydrogenase mutation diagram provided by the invention has higher conversion rate, stronger specificity and faster reaction speed.

The invention provides an application of a sunflower HaSnRK2 gene in regulating and controlling salt tolerance and drought tolerance of plants, and belongs to the technical field of plant genetic engineering. The invention provides an overexpression vector of a sunflower HaSnRK2 gene. The overexpression vector is applied to regulation and control of salt tolerance and drought tolerance of plants after genetic transformation is carried out on the plants by using a flower dipping method. Through overexpression of the sunflower HaSnRK2 gene, the salt stress tolerance and the drought stress tolerance of plants are remarkably enhanced, and damage caused by reactive oxygen free radicals induced by salt stress and drought stress can be effectively relieved.

The invention belongs to the field of genetic engineering and synthetic biology, and provides a novel cytochrome P450 enzyme derived from cephalotaxus plants, a nucleotide, a carrier and a host of the cytochrome P450 enzyme, and application of the cytochrome P450 enzyme in preparation of cephalotaxane diterpenoid compounds. The cytochrome P450 enzyme provided by the invention has multiple catalytic functions, can efficiently catalyze oxidation of C-5 site and C-19 site of cephalotaxene, lays a solid foundation for analysis of a cephalotaxane diterpene biosynthetic pathway, and provides a new method for obtaining important active molecules such as cephalotaxus hainanensis lactone and the like by adopting a synthetic biological means.

The invention relates to the technical field of plant genetic engineering, in particular to application of a rice OsERF141 protein and a coding gene thereof in improving low-temperature tolerance of plants. Experiments show that the low-temperature resistance of an OsERF141 function deletion mutant strain is remarkably reduced, and the low-temperature resistance of an overexpressed OsERF141 transgenic strain is remarkably improved. The invention provides a new gene resource for the cold resistance problem of rice, and has important significance for the molecular mechanism research of low-temperature stress and the cultivation of new low-temperature-resistant varieties.