5556 results about "Genetic engineering" patented technology

The present invention relates to the fields of genetic engineering and plant genetic breeding, and specifically to the application of wheat TaPND gene. In the present invention, by taking gene TaPND as a target, designing a sgRNA sequence based on CRISPR / Cas9, connecting a DNA fragment containing the encoding sgRNA sequence to a vector carrying CRISPR / Cas, a plant binary expression vector capable of knocking out the TaPND gene in wheat is constructed; by genetically transforming wheat, offspring with TaPND gene knockout are obtained, and wheat mutants with early flowering time are obtained. Experiments have confirmed that transgenic wheat with TaPND gene knockout achieves the advancement of wheat flowering time. The technical scheme of the present invention provides a practical method for realizing rapid wheat breeding using genetic engineering technology, which has important breeding application value and broad market application prospects.

The invention relates to the field of rice gene engineering, and particularly provides OsSULTR2; the invention also discloses application of the 2 protein and the coding gene thereof in regulating and controlling the salt tolerance of rice. The protein meets the following conditions: B1) a protein with an amino acid sequence of SEQ ID NO.1; and B2) a fusion protein with the same function obtained by connecting a tag to the N end and / or C end of B1). The method is used for detecting OsSULTR2; the salt stress phenotype identification in the seedling stage is carried out on the transgenic rice with the gene knockout 1, 2, and the result shows that when the gene segment is deleted, the salt stress tolerance of the rice is improved, and the function and the application way of the gene are proved. Therefore, the OsSULTR2 of the present invention; the 2 protein and the coding gene thereof can regulate and control the salt tolerance of rice, and have important significance for cultivating salt-tolerant transgenic rice.

The invention relates to a saccharomyces cerevisiae strain for efficiently synthesizing cordycepin as well as a construction method and application of the saccharomyces cerevisiae strain, and belongs to the technical field of microbial genetic engineering. The construction method provided by the invention comprises the following steps: carrying out multi-copy genome integrated expression on heterologous codon optimized 2 '-carbonyl-3'-deoxyadenosine reductase genes and 3 '-adenosine monophosphate phosphohydrolase genes, so as to increase the dosage of key enzymes; and supplementing the defective gene of the engineering strain to obtain the saccharomyces cerevisiae strain B5U3 with high yield of cordycepin. According to the obtained saccharomyces cerevisiae strain for efficiently synthesizing the cordycepin, the cordycepin yield reaches 2.92 g / L after shake flask fermentation, the cordycepin yield reaches 7.5 g / L after fermentation in a fermentation tank, and a foundation is laid for safer and more reliable industrial production of the cordycepin.

The invention discloses polyketide synthase or a polyketide synthase mutant and application thereof, and belongs to the technical field of gene engineering. The polyketide synthase is AspiPKS4, AspiPKS5, AspiPKS6 and AspiPKS7 from cyathea spinulosa, FhPKS2, FhPKS3, FhPKS4, FhPKS5 and FhPKS6 from ficus himalayana, and CrPKS from floating fern. The mutant is obtained by carrying out single point mutation on the second amino acid of AspiPKS7 or deleting an N-terminal redundant sequence. The polyketide synthase or the mutant is used for replacing NnHisspS in an original FBP luminescence system, and the biological self-luminescence level can be remarkably improved. The invention further provides a simplified and improved FBP system which comprises the polyketide synthase or the polyketide synthase mutant, CPH, H3H and Luz, and the application convenience and the application luminescence capacity of the FBP system are improved.

The invention relates to the field of gene engineering, and discloses a D-3-phosphoglycerate dehydrogenase mutant, a coding gene thereof and application of the D-3-phosphoglycerate dehydrogenase mutant and the coding gene. Compared with a protein with an amino acid sequence as shown in SEQ ID NO: 1, the mutant comprises mutation at the 131 site and / or the 285 site. The mutant can specifically catalyze a substrate 3-phosphoglycerate to synthesize 3-phosphate-hydroxypyruvic acid, can effectively promote synthesis of L-serine in a metabolic pathway of microorganisms, improves the yield and conversion rate of the L-serine, and reduces the yield of byproducts at the same time.

The invention discloses a tea tree CsU3 gene promoter and application thereof, and belongs to the technical field of plant biotechnology and gene engineering. Tea tree RNA polymerase III type promoters-tea tree endogenous U3 gene promoters CsU3a, CsU3b, CsU3c and CsU3d are successfully cloned for the first time, a tea tree U3 gene promoter activity detection carrier is successfully constructed, the transcriptional activities of the four CsU3 promoters are compared through instantaneous tobacco leaf conversion, GUS dyeing and GUS enzyme activity determination, and the CsU3 gene promoter activity detection carrier can be used for detecting the activity of the tea tree U3 gene promoter. CsU3a and CsU6b promoters with strong transcriptional activity are selected to be used for constructing a tea tree gene editing carrier, the feasibility of the promoter applied to a tea tree CRISPR / Cas9 gene editing system is verified in a tea tree protoplast, and CRISPR / Cas9 mediated tea tree genome targeted editing is realized.

The invention belongs to the technical field of gene engineering, and particularly relates to application of a ZmDIR11 gene in regulating and controlling salt stress resistance of corn. After salt stress treatment, it is found that the ZmDIR11 gene mutant corn plant growth vigor and material accumulation are significantly lower than those of a wild type B73 material, an oxidative metabolism regulation path, proline accumulation, chlorophyll synthesis, lignan accumulation and a Na < + > / K < + > transport pathway are inhibited, and related metabolic indexes are significantly lower than those of the wild type B73 material. It is shown that the ZmDIR11 gene positively regulates and controls the salt-resistant metabolic pathway of the corn in the corn, and the salt tolerance of the corn is reduced after the ZmDIR11 gene is mutated. The research provides a new gene resource for corn salt-tolerant breeding by utilizing gene engineering.

The invention belongs to the field of gene engineering, and particularly relates to application of an XC3225 gene in increasing the yield of xanthomonas campestris pv. Campestris xanthan gum. The invention finds that the XC3225 gene has a negative regulation effect on the xanthan gum yield, and the xanthan gum yield is remarkably improved by inhibiting or knocking out the XC3225 gene in the genome of xanthomonas campestris pv. Campestris, so that a new gene resource is provided for xanthan gum preparation.

The invention belongs to the technical field of genetic engineering, and particularly relates to recombinant collagen based on engineering bacterium expression and a preparation method thereof, and the recombinant collagen is formed by connecting repetitive units derived from human III type collagen in series for three times; the invention also discloses a base sequence of the coding gene of the recombinant collagen, efficient expression and accurate folding of target protein are realized through genetic modification of engineering bacteria, a light-operated dynamic folding process and an orthogonal purification strategy, and cascade purification of a His6-SUMO tag and an intein-CBD module is utilized to obtain the recombinant collagen. The product purity and the native conformation retention rate are remarkably improved, and the technical problems that hydroxyproline depends on exogenous addition, the folding controllability is poor and the purification efficiency is low in a traditional process are solved.

The invention discloses a PgEGY3 gene for improving cold resistance of pennisetum alopecuroides and increasing fresh weight of leaves and application of the PgEGY3 gene, and relates to the technical field of genetic engineering, and the nucleotide sequence of the PgEGY3 gene of the pennisetum alopecuroides is shown as SEQ ID NO.1. The invention also discloses a protein coded by the Pennisetum alopecuroides PgEGY3 gene. The invention also discloses a recombinant expression vector and a recombinant host cell comprising the Pennisetum alopecuroides PgEGY3 gene. The invention also discloses application of the PgEGY3 gene, a recombinant expression vector and a recombinant host cell in improving cold resistance and biomass of pennisetum alopecuroides. It is verified that the Pennisetum alopecuroides PgEGY3 gene can improve the cold resistance of plants, and the growth state of the plants under cold stress is remarkably improved through overexpression of the PgEGY3 gene.

The invention relates to the field of gene engineering for enhancing the cold resistance of plants, and relates to a recombinant expression vector, a recombinant strain and a primer pair of a pennisetum purpureum PpMYB2 gene, a method for improving the cold resistance of the plants and application of related materials in improving the cold resistance of the plants. One purpose of the invention is to provide application of the Pennisetum purpureum PpMYB2 gene and related materials thereof in improving the cold resistance of the plant, the amino acid sequence of the encoding protein of the Pennisetum purpureum PpMYB2 gene is as shown in SEQ ID NO.1, and the plant is arabidopsis thaliana or Pennisetum purpureum. Under the background that extreme low temperature events caused by global climate change are increasingly frequent, the cold resistance enhancing technology provided by the invention has important significance on guaranteeing the stability of agricultural production and reducing loss caused by low temperature. Through overexpression of the gene PpMYB2, the adaptability of perennial pennisetum purpureum to low-temperature stress can be remarkably improved, so that the planting range of perennial pennisetum purpureum is expanded to regions with lower temperature, and the survival and growth performance in different ecological environments is enhanced.

The invention discloses a pennisetum purpureum gene PpbHLH148L and application thereof in improving the cold resistance of plants, and relates to the technical field of gene engineering. The nucleotide sequence of the gene is as shown in SEQ ID NO.5. After the gene is subjected to subcellular localization and expression condition analysis after cold stress, the gene is related to low-temperature tolerance. An overexpression vector of the gene is constructed, Agrobacterium tumefaciens is transferred, then Arabidopsis thaliana infection is carried out, cold tolerance determination is carried out on overexpressed Arabidopsis thaliana, phenotypic difference and physiological index difference between an overexpressed strain after cold treatment and a wild type are observed, and it is found that the gene can significantly improve the low-temperature stress tolerance of Arabidopsis thaliana. And in the low-temperature treatment of the pennisetum alopecuroides transgenic callus, the transgenic callus also shows higher tolerance than the wild callus at low temperature. The pennisetum purpureum gene PpbHLH148L can be used for low-temperature tolerance modification of plants, and has an application prospect in creation of low-temperature-resistant plant materials.

The invention belongs to the technical field of genetic engineering, and particularly relates to an antibacterial peptide with antibacterial and antiviral effects and application and a product thereof. The amino acid sequence of the antibacterial peptide provided by the invention has the following general formula: X1GX2FX3FX4KKFX5LFX6X7SKVLK, x1 is M or has no amino acid; x2 is one of R, G and H; x3 is one of KS, KR, HR and HH; x4 is R or L; x5 is one of KK, HH and HK; x6 is RR or KK; and X7 is I or L. The antibacterial peptide disclosed by the invention has a relatively good antibacterial effect, and shows antiviral activity on HPV16 (Human Papilloma Virus 16) and HPV18 (Human Papilloma Virus 18). Good application prospects are shown in the preparation of antibacterial or antiviral products.

The invention provides a recombinant keratin polypeptide as well as a preparation method and application thereof, and relates to the technical field of genetic engineering. The amino acid sequence of the recombinant keratin polypeptide is as shown in any one of SEQ ID No. 1, SEQ ID No. 4, SEQ ID No. 7 and SEQ ID No. 10. The recombinant keratin polypeptide has a good soothing effect and the effects of stimulating skin angiogenesis and improving local blood circulation, and can be widely applied to the industries of medicines, cosmetics, skin care products and the like.

A resistance gene TaCAT to FCR of wheat and an application thereof are provided, which relates to the field of biological genetic engineering technologies. The nucleotide sequence of a genome where the TaCAT gene is located, the nucleotide sequence of a cDNA, the nucleotide sequence of a CDS and a coded protein sequence are respectively as shown in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4. An InDel marker is developed for identifying different allelotypes of the TaCAT gene. An application of the TaCAT gene for preparation of transgenic plant cells and plant breeding is provided. The molecular genetic basis of the resistance to FCR is disclosed, an important role is played in improving the resistance to FCR by utilizing a genetic engineering technology, and a new way is provided for breeding a new wheat variety with disease-resistance and safety.

The invention provides cat serum albumin and application thereof, and relates to the technical field of protein engineering. According to the invention, mutation is carried out on the cat serum albumin for the first time to obtain a beneficial mutation site, so that the cat serum albumin mutant which can be highly expressed in a pichia pastoris system is obtained. In addition, the cat serum albumin mutant I31L also has the effect of treating ascites due to cirrhosis, and the treatment effect is obviously higher than that in the prior art. A new biological material is provided for application of the cat serum albumin, and a foundation is laid for genetic engineering modification of the cat serum albumin.

The invention provides a recombinant vector for expressing LPL, a recombinant bacterium and an LPL production method, and belongs to the technical field of gene engineering. The invention provides a recombinant expression vector of Pseudomonas aeruginosa lipoprotein lipase LPL, wherein an LPL coding gene is derived from Pseudomonas aeruginosa. The recombinant expression vector is introduced into a pseudomonas aeruginosa PAO1 cell, an expression strain is successfully constructed, the expression quantity of target protein is improved, efficient expression of LPL in PAO1 is achieved, and the produced LPL is high in enzyme activity and wider in application range.

The specification provides methods for introducing a desired genetic change in a nucleotide sequence using a double-strand break (DSB)-inducing genome editing system, the method comprising: identifying one or more available cut sites in a nucleotide sequence; analyzing the nucleotide sequence and available cut sites with a computational model to identify the optimal cut site for introducing the desired genetic change into the nucleotide sequence; and contacting the nucleotide sequence with a DSB-inducing genome editing system, thereby introducing the desired genetic change in the nucleotide sequence at the cut site.

The invention belongs to the technical field of genetic engineering, and discloses a phytosusceptibility-related TaCNGC13 gene, a protein and application of the phytosusceptibility-related TaCNGC13 gene and the protein. The phytosusceptible related TaCNGC13 gene is derived from a wheat variety water source 11, the nucleotide sequence of the gene is shown as SEQ ID NO: 1, and the amino acid sequence of the gene encoding protein is shown as SEQ ID NO: 2. The plant-susceptibility-related TaCNGC13 gene is induced to express by the puccinia striiformis, the expression of the plant-susceptibility-related TaCNGC13 gene in wheat is stably silenced, and the resistance of the obtained transgenic wheat to a plurality of physiological races of the puccinia striiformis is higher than that of wild wheat. The invention provides a novel germplasm material for green control of wheat stripe rust and cultivation of wheat stripe rust resistant varieties.

The invention discloses a method for traceless gene editing of gram negative bacteria, a carrier and application of the carrier, and belongs to the technical field of microbial genetic engineering. The invention provides a vector for performing traceless gene editing on bordetella pertussis. The vector simultaneously comprises an antibiotic positive screening gene expression box and a sacB and galk negative screening gene expression box. According to the method, the sacB gene and the galk genome are synthesized into the double-reverse-screening system for the first time, and compared with a single-reverse-screening system, the reverse-screening efficiency is remarkably improved, and the dosage of a reverse-screening reagent is remarkably reduced. The invention provides a novel gene editing method for non-mode bacteria such as pertussis bordetella and the like, and has a good application prospect.

The invention discloses an application of a medicago sativa MsDGK5 gene and a protein coded by the medicago sativa MsDGK5 gene in salt stress resistance of plants, and belongs to the technical field of plant genetic engineering. The nucleotide sequence of the medicago sativa MsDGK5 gene provided by the invention is as shown in SEQ ID NO. 1. According to the application, MsDGK5 gene overexpressed arabidopsis thaliana and medicago sativa transformation plants are constructed, growth and development indexes and physiological and biochemical parameters of the plants under salt stress are systematically measured, and it is proved that overexpression of the MsDGK5 gene can significantly improve the salt tolerance of the transgenic plants. Based on the experimental results, the key regulation effect of the MsDGK5 gene in plant salt tolerance is defined for the first time, an important gene resource is provided for crop salt tolerance genetic improvement, and a new technical approach is also provided for coping with the increasingly aggravated soil salinization problem.

The invention belongs to the field of gene engineering, and relates to a DNA binding protein based on TraC effect protein, a fusion protein with a cytosine / adenine base editing function, an editing system and nucleic acid molecules or constructs for encoding the DNA binding protein and the fusion protein. The invention also relates to complexes and kits for gene editing comprising the fusion proteins of the invention or the nucleic acid molecules encoding them. The base editing tool disclosed by the invention can realize efficient, accurate and safe base editing in cells of various eukaryotes such as animals and plants. The unique characteristics of the base editing tool enrich the types of existing base editing tools, and the base editing tool has wide application prospects.

The invention provides an IbFLZ9 gene and application thereof in regulation and control of growth of sweet potato vines, and belongs to the technical field of gene engineering and sweet potato breeding. The invention provides an IbFLZ9 gene for regulating and controlling growth of sweet potato vines. The nucleotide sequence of the IbFLZ9 gene is shown as SEQ ID NO. 1. Growth of sweet potato vines can be remarkably inhibited by overexpressing the IbFLZ9 gene, and growth of sweet potato vines can be remarkably promoted by inhibiting expression of the IbFLZ9 gene through a gene knockout technology. A new sweet potato variety which is moderate in vine length and convenient to mechanically harvest can be created by regulating and controlling the expression abundance of the IbFLZ9 gene, and a new gene resource and a new breeding technology are provided for improving the characters of the overground part of the sweet potato.

The invention relates to the technical field of microbial genetic engineering, in particular to a method for improving cordycepin synthesized by saccharomyces cerevisiae and saccharomyces cerevisiae engineering bacteria. According to the invention, codon-optimized 2 '-carbonyl-3'-deoxyadenosine reductase gene and 3 '-adenosine monophosphate phosphohydrolase gene from cordyceps militaris are expressed by using plasmids, and then the plasmids are transformed into host cells to obtain the saccharomyces cerevisiae engineering bacteria. And inoculating a seed solution obtained by seed culture of the activated saccharomyces cerevisiae engineering bacteria into a fermentation culture medium containing a metabolic effector, and carrying out fermentation culture to synthesize cordycepin. Metabolic effectors (Cu < 2 + >, Fe < 2 + >, Mg < 2 + >, Zn < 2 + >, citric acid, cysteine, aspartic acid, glycine, VB1 and tea polyphenol) are added into a fermentation culture medium strain, and the variety and concentration of the metabolic effectors are optimized, so that the yield and synthesis efficiency of cordycepin are remarkably improved. By using the combination of the strain obtained by the invention and the optimal effector, the yield of cordycepin in a fermentation tank reaches 2.9 g / L.

The invention belongs to the field of gene engineering, and relates to a rice BRW1 gene and application thereof. The cDNA (complementary deoxyribonucleic acid) sequence of the rice BRW1 gene is shown as SEQ ID NO.1, and the amino acid sequence of the RNA binding protein coded by the rice BRW1 gene is shown as SEQ ID NO.2. The invention discloses the rice RNA binding protein gene BRW1 and the protein coded by the rice RNA binding protein gene BRW1. The rice BRW1 gene controls the grain weight of grains, the thousand grain weight of a knockout mutant of the gene is remarkably increased, and meanwhile the grain length and the grain width are increased. By mutating the BRW1 gene in the invention, the grain weight of rice grains can be increased, which indicates that the genetic improvement of the grain weight and grain shape of rice is facilitated by using the gene.

Provided is the use of overexpression of the auxin transporter PtoPIN1a gene in increasing the biomass of Populus tomentosa. The nucleotide sequence of the auxin transporter PtoPIN1a gene is as shown in SEQ ID No. 1. By means of using a genetic engineering method, wild-type Populus tomentosa is transformed by means of Agrobacterium tumefaciens infection to obtain a tissue-specific PtoPIN1a overexpressing transgenic plant, and subsequently, a plant with a relatively high expression level is selected, by means of positive identification, for phenotype observation. Results show that compared with the wild type in the same period, the PtoPIN1a transgenic plant has an increased plant height, a thicker stem and a significantly increased aboveground biomass. By means of the provided technical cultivation method, a high-biomass high-quality poplar variety is cultivated. The method is of great significance for future molecular breeding to cultivate excellent poplar varieties.

The invention discloses a hybridoma cell strain, a monoclonal antibody, a linear epitope antigen and application thereof, and relates to the field of genetic engineering, in particular to a hybridoma cell strain, a monoclonal antibody, a linear epitope antigen and application thereof. The preservation number of the hybridoma cell strain P27-A1 is CGMCC (China General Microbiological Culture Collection Center) No. 46352. The hybridoma cell strain P27-A1 is prepared from feline leukemia virus p27, and a monoclonal antibody secreted by the hybridoma cell strain P27-A1 recognizes a linear epitope antigen of feline leukemia virus p27 protein; the kit for the feline leukemia virus p27 comprises the monoclonal antibody A1 secreted by the hybridoma cell strain P27-A1. The monoclonal antibody secreted by the hybridoma cell strain can greatly improve the sensitivity of the p27 protein for detecting feline leukemia virus. The invention provides the monoclonal antibody secreted by the hybridoma cell strain for identification, and the monoclonal antibody can be used for preparing a diagnostic kit such as a colloidal gold test strip.

The invention discloses application of a PagBAG8 gene in regulation and control of poplar growth and development, and belongs to the technical field of forestry biological genetic engineering. According to the application of the PagBAG8 gene in regulating and controlling growth and development of the poplar, the nucleotide sequence of the PagBAG8 gene is shown as SEQ ID NO: 1, and the method for cultivating the transgenic poplar through the PagBAG8 gene comprises the following steps that the PagBAG8 gene of the poplar is cloned; the PagBAG8 gene is connected to a vector, and an overexpression vector is obtained; transforming the obtained overexpression vector into agrobacterium to obtain an agrobacterium liquid; and infecting poplar leaves with the obtained agrobacterium liquid to obtain a PagBAG8 gene overexpression plant. The invention provides a new choice for regulating and controlling the growth and development of the poplar, and provides a potential target for molecular breeding improvement of the properties of the poplar.

The invention discloses a light-driven synthetic flora and application thereof. The light-driven synthetic flora comprises phototrophic bacteria and heterotrophic bacteria, the phototrophic bacteria are genetically modified synechococcus slender, and the gene modification comprises construction and expression of heterologous sucrose permease, glyceraldehyde dehydrogenase and sucrose synthase in the synechococcus slender; and the heterotrophic bacteria are the P.adami. According to the light-driven synthetic flora disclosed by the invention, carbon dioxide is converted into cane sugar through photosynthesis of phototrophic bacteria, and the cane sugar is secreted out of cells to provide a carbon source for heterotrophic bacteria; carrying out cell growth and polyhydroxyalkanoate synthesis on the heterotrophic bacteria by using cane sugar; therefore, the light-driven synthetic flora provided by the invention can realize direct conversion of carbon dioxide into polyhydroxyalkanoate without additional carbon sources, has the advantage of environmental friendliness, and provides a new economical and feasible scheme and approach for production of polyhydroxyalkanoate.

The invention belongs to the technical field of gene engineering, and relates to a cyperus esculentus CeMYB154 gene capable of improving salt tolerance and active oxygen scavenging capacity and application of the cyperus esculentus CeMYB154 gene. The Cyperus esculentus CeMYB154 gene belongs to an MYB transcription factor family, the Cyperus esculentus CeMYB154 gene is screened and transformed into Arabidopsis thaliana through transcriptome sequencing, bioinformatics analysis, gene cloning, overexpression vector construction and recombinant strain overexpression, and analysis finds that under salt stress, compared with a wild type, the root length of a plant with the overexpressed CeMYB154 gene is obviously increased, the growth vigor is better, and the growth rate of the plant with the overexpressed CeMYB154 gene is higher. According to the present invention, the content of MDA and H2O2 is significantly reduced, and the activity of CAT, POD and SOD antioxidant enzymes is significantly enhanced, such that the overexpressed CeMYB154 gene can enhance the salt tolerance and the active oxygen scavenging ability of the plant, and the overexpressed CeMYB154 gene is proved to be the salt tolerance positive regulation factor; the CeMYB154 gene can be used in the field of salt-tolerant stress-resistant breeding of cyperus esculentus, and provides an important gene resource for salt-tolerant molecular breeding.