1305 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 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 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 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 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.

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 discloses engineered saccharomyces cerevisiae and application of the engineered saccharomyces cerevisiae in synthesis of 2-phenethyl alcohol, and belongs to the technical field of genetic engineering. According to the engineering saccharomyces cerevisiae, saccharomyces cerevisiae CICC 33068 is modified, and the strain is modified through strategies such as aldrin pathway enhancement, precursor enhancement, tolerance optimization and cofactor balance. The constructed engineered strain has good metabolic performance and stability, not only is the tolerance of the engineered strain to 2-phenethyl alcohol improved, but also the production capacity of the 2-phenethyl alcohol is remarkably improved. Therefore, the engineered saccharomyces cerevisiae disclosed by the invention has remarkable advantages in production of the 2-phenethyl alcohol, can provide technical guidance for industrial production of the 2-phenethyl alcohol, and has a good application prospect.

The invention belongs to the technical field of plant genetic engineering, and discloses application of an LcCBP60b-LcALY2 molecular module in regulation and control of growth and development and maturing rate of leymus chinensis. The invention discovers that the calmodulin binding protein coding gene LcCBP60b negatively regulates the growth and development and the maturing rate of the leymus chinensis for the first time, and a yeast double-impurity experiment discovers that the LcCBP60b interacts with the LcALY2 protein to regulate the related characters of the maturing rate of the leymus chinensis. The LcCBP60b negatively regulates the activity of the LcALY2 protein and inhibits the regulation effect of the LcALY2 coded THO protein complex 4B in the flowering and ear forming process of leymus chinensis. The LcCBP60b-LcALY2 molecular module verified by the invention provides a new perspective for deeply understanding a Leymus chinensis fruiting regulation mechanism, and provides a direct molecular target for directionally creating new varieties of high-fruiting-rate pasture and cereal crops.

The invention provides a genetically engineered bacterium for producing polydeoxyribonucleotide as well as a construction method and application of the genetically engineered bacterium. The genetically engineered bacterium comprises kluyveromyces marxianus for expressing a salmon DNA polymerase I gene. According to the invention, the edible probiotic Kluyveromyces marxianus is transformed through genetic engineering, efficient, safe and controllable production of PDRN is realized, the biological activity of PDRN is equivalent to that of PDRN from a natural source, and large-scale production and application of PDRN are facilitated.

The invention discloses a 5 'UTR library, a recombinant expression vector and a genetically engineered bacterium for improving the yield of folamin in pathogenic bacterium Badei and application of the 5' UTR library and the genetically engineered bacterium, and belongs to the technical field of microbial genetic engineering. The method comprises the following steps: designing an SD region based on a 16S rRNA (ribosomal Ribonucleic Acid) 3'end complementary sequence of a Buddei pathogenic bacterium XBD101 serving as an original strain, regulating and controlling the number of bases between the SD sequence and an initiation codon AUG and optimizing an UTR secondary structure to construct a library containing 14 UTR sequences, and replacing three sequences N4A, N7A and N18A with the strongest translation ability in the library to the UTR region of a folamin synthesis key gene fclC, so as to obtain the folamin peptide. And finally, the capacity upgrading of the folamin chassis strain is realized, and an important support is provided for the industrial production of agricultural antibiotics.

The invention relates to an STING gene knockout TIL capable of enhancing anti-tumor activity and a preparation method and application of the TIL, in particular to a genetically engineered immune cell, and an STING gene in the genetically engineered immune cell is silenced or down-regulated. The ferroptosis resistance of the immune cell provided by the invention is remarkably enhanced, the immune cell can be better infiltrated into a tumor microenvironment, and the immune cell has a remarkable anti-tumor effect.

The invention specifically discloses a salix cheilophila in-situ genetic transformation method independent of a tissue culture system, and belongs to the field of plant genetic engineering. According to the method, semi-lignified branches of salix cheilophila are taken as receptors, vacuumizing treatment is performed by utilizing agrobacterium rhizogenes infection liquid, hairy roots are directly induced under a water culture condition after short-term dark culture, and positive identification is performed through reporter genes. According to the method, tedious sterile tissue culture operation is completely avoided, the transformation period is shortened to about 22 days, and the technical threshold and the time cost are remarkably reduced. By adopting the system, the positive rate can reach 54.33% when the pBI121 vector is used for transformation, and the efficiency is remarkably improved compared with that of an existing method. The method provides an efficient and reliable technical platform for root system gene function research, expression analysis and genetic improvement of salix cheilophila and related species thereof, and has high application potential.

The invention relates to an application of a FaPDHE1alpha gene or an encoding protein thereof in prevention and control of strawberry gray mold, and belongs to the field of plant genetic engineering and the field of prevention and control of postharvest diseases of fruits and vegetables. The invention discloses a key positive regulation and control effect of FaPDHE1alpha in regulation and control of strawberry gray mold resistance. By constructing a 35S:: FaPDHE1alpha overexpression vector and a FaPDHE1alpha RNAi interference vector, according to transient gene overexpression and RNA interference silencing tests, it is verified that high expression of the FaPDHE1alpha gene can obviously enhance the resistance of strawberry fruits to botrytis cinerea, and silencing of the gene causes reduction of the disease resistance of the fruits. Mechanism studies show that after the FaPDHE1alpha gene is activated, mitochondrial energy metabolism and controlled ROS signal generation are promoted, antioxidant enzyme activity is enhanced, and defense related gene expression is induced, so that the disease resistance of fruits is improved. By means of functional genomics, the key function of the FaPDHE1alpha gene in prevention and control of gray mold after strawberry harvest is illuminated for the first time, and the FaPDHE1alpha gene can be used for screening disease-resistant breeding materials or developing biological prevention and control preparations for postharvest diseases and has important application value.

The invention belongs to the field of metabolic engineering, and discloses a genetically engineered bacterium for producing myo-inositol as well as a construction method and application of the genetically engineered bacterium. The genetically engineered bacterium takes escherichia coli as an original strain, and is obtained by performing the following gene editing on a genome of the escherichia coli: non-expressed lactose operon repressor protein, glucose phosphate dehydrogenase, acetokinase, glucose phosphate isomerase and glycerol repressor protein; and overexpressing inositol-1-phosphate synthase, inositol monophosphate, glucokinase, glucose permease and glycerol kinase. Compared with the prior art, the genetically engineered bacterium disclosed by the invention has remarkable advantages in the aspects of key enzyme expression rate, metabolism specificity, fermentation period and the like, and an efficient, stable, economical and feasible solution is provided for industrial production of myo-inositol.

The invention discloses nano-enzyme coating recombinant probiotics as well as a preparation method and application thereof, and belongs to the technical field of biotechnology and nano medicine. According to the invention, engineered Escherichia coli Nissle 1917 (ECN (DE3)) is taken as a host, and glutathione peroxidase (GPX) and glutathione reductase (GR) genes are over-expressed through a genetic engineering means, so that an efficient antioxidant enzyme system is constructed. And sequentially modifying chitosan and cerium dioxide nano-enzyme (CNP) on the surfaces of the thalli by using an electrostatic layer-by-layer self-assembly technology to form a protective coating. The nano-enzyme coating significantly improves the survival rate of the recombinant probiotics in severe environments such as simulated gastric acid (pH 2.5) and bile salt (0.3%), and enhances the ability of the recombinant probiotics to remove reactive oxygen species (ROS), including hydrogen peroxide, superoxide anions, hydroxyl free radicals and DPPH free radicals.

The application discloses an Escherichia coli probiotic with high beta-alanine yield and application thereof, and the recombinant genetically engineered bacterium takes a wild-type Escherichia coli probiotic as a starting strain and is subjected to one or more gene edits, including Trc promoter strengthening of a panD gene, an aspC gene, an sdhC gene and a ppc gene, overexpression of a panD gene from Bacillus subtilis, an aspB gene from Corynebacterium glutamicum, an aspA gene and a pyc gene from Corynebacterium glutamicum on a plasmid pTrc99a, and knockout of a cycA gene. 600 Compared with an existing reported Escherichia coli probiotic beta-alanine production strain, the OD of the recombinant genetically engineered bacterium is 4.4 times that of the existing reported strain, and the beta-alanine yield is 6 times that of the existing reported strain, so that the recombinant genetically engineered bacterium has great industrial application potential.

The invention relates to the field of synthetic biology, metabolic engineering and fermentation engineering, and discloses a genetically engineered bacterium and application thereof in production of alpha-ketoglutaric acid.The genetically engineered bacterium comprises the steps that the genetically engineered strain with escherichia coli BW25113 or corynebacterium glutamicum ATCC13032 as a host is constructed; overexpression type isocitrate dehydrogenase genes are integrated through chromosomes, alpha-ketoglutarate dehydrogenase complex genes are knocked out, NADPH invertase is overexpressed through plasmids, and exogenous AKG transporter protein is introduced, so that metabolic flux directional regulation and control and product efflux enhancement are realized; meanwhile, a staged fermentation process is adopted and comprises dynamic gradient adjustment of inorganic phosphorus concentration and two-stage pH control. According to the method, the AKG yield of 78.2 g / L can be obtained under the 5L tank scale, the sugar acid conversion rate reaches 0.68, and the method has the advantages of being high in yield, low in byproduct and controllable in cost. The obtained AKG is suitable for a plurality of fields such as medical intermediates, functional foods and cell culture additives, and has wide industrial application prospects.

This invention specifically relates to a de novo 7-DHC-synthesizing *Saccharomyces cerevisiae* genetically engineered strain, its construction method, and its applications, belonging to the field of biotechnology. Using *Saccharomyces cerevisiae* engineered strain SC9 as the starting strain, this invention utilizes multiple copies of tHMG1, DHCR24, and ERG1 via the TY transposon, multiple copies of ERG2, POS5, ERG7, ERG11, IDI1, and ERG3 via the TY transposon, and single copies of POS5, ERG3, and DHCR24 in the genome to increase the intracellular content of reducing coenzyme II and enhance the main pathway of 7-DHC synthesis. Single copies of ACL, ACS1, ERG10, and ERG19 in the genome enhance the utilization of ethanol in *Saccharomyces cerevisiae* and dynamically regulate the synthesis of 7-DHC. The *Saccharomyces cerevisiae* genetically engineered strain constructed by this invention exhibits high product accumulation, laying the foundation for replacing traditional multi-step chemical synthesis routes and realizing industrial production.

The invention discloses a limonium bicolor gene LbZFP8 and application thereof, and relates to the technical field of biological genetic engineering. The invention relates to a limonium bicolor gene LbZFP8 and application thereof. The limonium bicolor gene LbZFP8 is a gene for coding a protein composed of an amino acid sequence as shown in SEQ ID NO: 1; the biological material is recombinant DNA, an expression cassette, a transposon, a plasmid vector, a virus vector or engineering bacteria. The gene provided by the invention can significantly improve the salt tolerance and oxidation resistance of a target plant by transforming plant cells. Experiments show that under the salt stress condition, the survival rate of the LbZFP8 overexpressed arabidopsis thaliana plant is increased by about 45% compared with that of a control group. Meanwhile, the compound plays a key role in regulating and controlling an active oxygen scavenging system, and the activity of SOD and POD enzymes can be effectively enhanced. The technology provides an important gene resource for cultivating new varieties of crops with strong stress resistance, and has a wide application prospect.

The invention belongs to the technical field of bioengineering, and discloses a genetic engineering strain for enhancing schizochytrium limacinum DHA oil production based on carbon flux guidance and fatty acid transport, a method and application thereof. The gene engineering strain is obtained by introducing an optimized coding gene ACLY and / or Fat1p coding gene related to guiding carbon flux and fatty acid transport into a schizochytrium limacinum body; wherein the gene sequence of the gene ACLY is SEQ ID No. 9, and the gene sequence of the gene Fat1p is SEQ ID No. 10. According to the invention, a schizochytrium limacinum engineering bacterium for enhancing DHA oil production based on carbon flux guidance and fatty acid transfer is constructed by means of metabolic engineering. The schizochytrium limacinum engineering bacterium is combined with fatty acid transfer based on carbon flux guiding, can be used for grease synthesis by guiding carbon flux, and can prevent grease from being degraded through fatty acid transfer.

The invention discloses a genetically engineered bacterium for biosynthesis of gastrodin as well as a construction method and application of the genetically engineered bacterium. According to the genetically engineered bacterium, through overexpression of sucrose permease CscB derived from escherichia coli W, sucrose phosphorylase Basp of bifidobacterium adolescentis, uridine triphosphoryl-glucose-1-phosphate uridine acyl transferase UgpA of bifidobacterium bifidum and UDP-glycosyl transferase RsUGT of hedyotis diffusa, an exogenous path for synthesizing UDP-glucose from sucrose is constructed in a cell, and the UDP-glucose is synthesized into UDP-glucose. And the glycosylation capability of the gastrodin precursor on hydroxybenzyl alcohol is enhanced. The invention also provides a construction method of the genetically engineered bacterium and a method for synthesizing gastrodin through whole-cell catalysis by using the strain and taking p-hydroxybenzyl alcohol and cane sugar as substrates. The genetically engineered bacterium is high in gastrodin synthesis efficiency, the molar conversion rate can reach 97.8%, the highest yield reaches 3.61 g / L, and a new green manufacturing scheme is provided for solving the problems that a traditional gastrodin production method is low in efficiency, high in cost, large in pollution and the like.

The present application relates to expression constructs and methods for genetically engineering methylotrophic yeasts. The present invention provides methods and materials for genetically engineering methylotrophic yeasts.

Therefore, this disclosure provides a genetically engineered mesenchymal stem cell (MSC) population, including an expression vector containing the Akt or HGF gene and the PD-L1 gene. It also provides a method for synergistically improving the survival status and immunomodulatory capacity of mesenchymal stem cells or enhancing their proliferation, including transfecting mesenchymal stem cells with the Akt or HGF gene and the PD-L1 gene; and a method for preventing, improving, and / or treating ischemic conditions, enhancing neurogenesis, or reducing neuronal death, including administering an effective amount of the genetically engineered mesenchymal stem cell population of this disclosure to individuals in need.

The present application belongs to the field of cell therapy and immunotherapy, and provides a genetically engineered human trophoblast, a preparation method and application thereof. The human trophoblast takes K562 cells as starting cells, and stably expresses membrane-bound interleukin 21, CD137 ligand and Delta-like ligand 1 after genetic engineering. The constructed K562 three-factor trophoblast can significantly improve the expansion efficiency, activation state and functional stability of NK cells and γδT cells. The synergistic mechanism includes enhancing the proliferation, cytotoxicity and stemness maintenance of NK cells and γδT cells through STAT3, NF-κB and Notch signaling pathways, respectively. The human trophoblast has the advantages of good expression stability, significant functional enhancement, and high activity after freezing and recovery.

The present application relates to the field of biological genetic engineering technology, in particular to the application of silencing miR529b in the positive regulation of rice resistance to brown planthopper. The silencing vector of miR529b is obtained by short tandem target mimic (STTM) technology, which silences miR529b in the receptor rice, and obtains the transgenic plant STTM-miR529b with silenced miR529b. The T2 generation homozygous transgenic plants are identified for brown planthopper resistance by seedling group method. Compared with the transgenic receptor parent Nipponbare (control group, marked as NIP, same below), STTM-miR529b is more resistant to pests, which confirms that miR529b is related to the resistance of rice to brown planthopper, and can be used for creating new germplasm of rice resistance to brown planthopper.

The invention belongs to the technical field of microbial pharmacy, and discloses a polyketone macrolide compound generated by genetically engineered streptomyces as well as a preparation method and application thereof. The polyketone macrolide compound disclosed by the invention is a cinnamyl streptomycin derivative, has cell proliferation inhibition activity better than that of a natural product Cinnamomycin A-D, can exert anti-tumor activity by inhibiting activity of human exonucleotide pyrophosphatase / phosphodiesterase 1 (ENPP1) and activating an inherent immune pathway, can be prepared into a medicine or a medicinal composition, and can be used for preparing medicines or medicinal compositions. The compound is used for treating tumors and related diseases.