2842 results about "Wild type" patented technology

The invention belongs to the technical field of gene engineering, and particularly relates to applications of OpgG and a mutant thereof in production of human milk oligosaccharide. The invention provides a dextran biosynthetic protein OpgG mutant, and the OpgG mutant is obtained by mutating the 443rd amino acid from threonine to proline on the basis of a wild type dextran biosynthetic protein OpgG as shown in SEQ ID NO.1. The invention also provides a preparation method of the dextran biosynthetic protein OpgG mutant. An opgG gene and / or a T443P mutant coding gene are / is introduced into a human milk oligosaccharide production strain and applied to production of human milk oligosaccharide, and the yield of 3 '-SL, 6'-SL, LNT II, LNnT and LNT is greatly increased.

The invention discloses a high temperature resistant xanthan gum lyase mutant and a preparation method thereof, wild type xanthan gum lyase comes from Paenibacillus anensis, site-directed mutagenesis is carried out on the wild type xanthan gum lyase to obtain the high temperature resistant xanthan gum lyase mutant, and the mutant comprises at least one of N66V, R292A and S676V. The enzymatic activity of the xanthan gum lyase mutant treated in a buffer solution with the pH value of 5.0 at the temperature of 65-85 DEG C for 2 h is remarkably improved compared with that of an original enzyme, and the enzymatic activity of N66V / R292A / S676V is improved by 3.8 times compared with that of the original enzyme at the temperature of 85 DEG C.

The invention discloses a hemoglobin-resistant Taq DNA polymerase mutant and a construction method thereof, and relates to the field of biology, and the hemoglobin-resistant Taq DNA polymerase mutant is characterized in that a nucleotide sequence for coding the Taq DNA polymerase is shown as SEQ ID NO.1, and the mutant comprises at least one mutation or all mutations selected from S623D and E721A sites. On the basis of natural Taq DNA polymerase, the molecular structure of the Taq DNA polymerase is modified through rational design and combination with a site-specific mutagenesis biotechnology, so that the Taq DNA polymerase mutant is more suitable for PCR amplification of samples containing hemoglobin and the like than a wild type, and the result judgment accuracy of a sample containing a hemoglobin inhibitor is more facilitated.

The invention includes a mutant T7 DNA ligase or a biologically active fragment thereof, which has greater activity than wild type T7 DNA ligase. The mutant T7 DNA ligase, or the biologically active fragment, has one or more substitutions differing from the wild type, as described more fully in the Summary. The preferred mutant T7 DNA ligase has at least one of the following mutations: E63K (SEQ ID NO:4), K73E (SEQ ID NO:6), K137E (SEQ ID NO:7), K174E (SEQ ID NO:9), E182K (SEQ ID NO:11), K210E (SEQ ID NO: 13), E243K (SEQ ID NO:15), D245R (SEQ ID NO: 17), E268K (SEQ ID NO:19), E272K (SEQ ID NO:21), E289K (SEQ ID NO:23), K295E (SEQ ID NO:25) and D336R (SEQ ID NO:27).

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 discloses application of a TaLBD30 protein and a coding gene thereof in regulating and controlling a wheat plant type. The invention belongs to the technical field of biology, and particularly relates to application of TaLBD30 protein and a coding gene thereof to regulation and control of wheat plant types. The protein is any one of the following proteins: A1) a protein with an amino acid sequence as shown in SEQ ID No: 1; a2) a protein which is obtained by substitution and / or deletion and / or addition of amino acid residues on the protein of A1), has 80% or more of identity with the protein of A1) and has the same function as the protein of A1); a3) a fusion protein obtained by connecting a protein tag to the N terminal or / and the C terminal of A1) or A2). TaLBD30 is a transcription factor with transcription inhibition activity, and compared with a wild type, the TaLBD30 gene overexpression shows that after TaLBD30 overexpression, the wheat plant height and ear length are remarkably reduced.

The invention includes a mutant Taq polymerase, which can significantly extend and amplify a target sequence where the extension conditions are time limited to as little as one second. The mutant Taq polymerase, or a biologically active fragment thereof, has one or more substitutions differing from the wild type as shown in Table I.

The invention discloses an application of a rape BnC05ERF378 gene in improving the waterlogging resistance of a plant. The nucleotide sequence of the rape BnC05ERF378 gene is as shown in SEQ ID NO. 1. According to the invention, a brand new stain-resistant gene BnC05ERF378 is cloned from rape for the first time, and is transferred into arabidopsis thaliana to construct a pure line plant and complete stain resistance identification. Experimental results show that the growth state of the transgenic BnC05ERF378 gene arabidopsis thaliana is remarkably superior to that of a wild type after waterlogging treatment, the transgenic BnC05ERF378 gene arabidopsis thaliana can normally bolt, the survival rate is increased from 12.5% to 37.5%, it is fully proved that the gene can enhance the survivability of the plant in a waterlogging environment by regulating the response mechanism of the plant to waterlogging stress, and the survival rate of the plant is increased from 12.5% to 37.5%. And a key guarantee is provided for stable growth of plants under high-humidity, waterflooding and other adverse conditions. According to the discovery and application of the waterlogging-resistant gene BnC05ERF378 disclosed by the invention, a clear functional gene target is provided for improvement of waterlogging resistance of crops. By constructing an expression vector containing the gene and combining mature technologies such as agrobacterium transformation, the gene can be stably introduced into a target plant, and a pure transgenic plant is obtained.

The invention belongs to the technical field of characterization of target samples, and provides PANX3 biopore protein and application of a mutant and single molecule detection thereof. The PANX3 biopore protein is composed of seven identical subunits, the overall channel is funnel-shaped, and the amino acid sequence is shown as SEQ ID NO: 1. After one or more amino acids of the wild type PANX3 biopore protein are mutated into common amino acids except original amino acids, compared with the wild type PANX3 biopore protein, the mutant can be normally expressed and has stable properties, and the current property and the current signal of the mutant are obviously improved.

The invention discloses a gene PtoMYB20 for regulating and controlling salt tolerance and drought resistance of poplars. A nucleotide sequence of the gene comprises a sequence as shown in SEQ ID NO. 1. The gene interference fragment is introduced into a plant to obtain a silent transgenic plant, under the conditions of salt stress, drought stress and salt-drought composite stress, the H2O2 and MDA contents of the silent transgenic plant are lower than those of a wild plant, and the SOD and POD activities of the silent transgenic plant are higher than those of the wild plant, so that the damage degree of the plant is effectively reduced; the method is of great significance in improving salt tolerance and drought resistance of the poplar and relieving damage of adversity stress to forestry yield. The invention also discloses a breeding method of the transgenic poplar with strong salt tolerance and drought resistance, provides a new poplar germplasm which not only improves the stress resistance of the poplar but also does not influence the wood yield, and lays a theoretical and gene resource foundation for the stress-resistant molecular breeding of the poplar.

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 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.

The present invention belongs to the field of nucleic acid editing, and particularly the technical field of clustered regularly interspaced short palindromic repeats (CRISPRs). Specifically, provided in the present invention is a Cas mutant protein having an improved activity and an improved editing efficiency. Compared with a wild-type parental Cas protein, the Cas mutant protein of the present invention has significantly improved activity, and has a wide application prospect.

The invention provides an RNA polymerase variant and application thereof, and relates to the technical field of biology, the enzyme specific activity of the variant is obviously improved compared with that of a wild type, and the addition amount of polymerase is reduced, so that the cost is saved. Meanwhile, the mutant also effectively reduces the generation of incomplete fragment impurities and improves the integrity. In addition, the invention further provides a method for generating RNA through in-vitro transcription.

The present invention relates to a method for constructing an eight-gene-edited xenogeneic organ transplantation donor pig, belonging to the field of animal biotechnology. In the wild-type porcine fetal fibroblast cell line, the GGTA1, β4GalNT2, and CMAH genes are knocked out by using the CRISPR / Cas9 gene editing technology, and the humanized genes of hCD39, hCD46, hCD55, hCD59, and hTBM are transfected. Combining with somatic cell cloning technology, GTKO / β4GalNT2KO / CMAHKO / hCD39 / hCD46 / hCD55 / hCD59 / hTBM eight-gene-edited cloned pigs are constructed. Further, through genotype, mRNA, protein expression identification and functional analysis, eight-gene-edited xenogeneic organ transplantation donor pigs are obtained. The present invention solves the technical problems of high production difficulty, low efficiency, and low survival rate of donor pigs for multi-gene-edited xenogeneic organ transplantation, and maximally solves the common problems of immune rejection reaction and complement dysregulation faced during xenogeneic organ transplantation, laying a foundation for more targeted development of donor pigs suitable for different tissue and organ xenotransplantation, and having important value for promoting the clinical transformation of xenogeneic organ transplantation.

The invention provides an EGFR wild-type lung adenocarcinoma prognosis risk assessment method based on multi-omics and machine learning, and the method comprises the steps: obtaining multi-omics and clinical data of lung adenocarcinoma, obtaining a data set, and carrying out the multi-omics consensus clustering, and obtaining a molecular typing result; high-risk subtype specific candidate genes are identified, a candidate prognosis gene set is obtained, multi-algorithm machine learning comparison optimization is carried out, and a modeling strategy is obtained; performing feature screening and model training to obtain a multi-omics feature model so as to calculate an individual risk score of the to-be-tested sample; the individual risk score and the clinical staging information are utilized to obtain a clinical column diagram and a survival prediction result, then the flow of the multi-omics feature model, the individual risk score and the survival result is Web to obtain a clinical system, and a lung adenocarcinoma prognosis risk assessment result is output. The invention can realize an objective, accurate, generalizable and multifunctional prognosis evaluation and treatment guidance tool, and has important clinical application value and wide industrialization prospect.

The invention provides an RPA-CRISPR-Cas12a system-based detection reagent for the drug resistance of a chilo suppressalis organophosphorus insecticide, a detection method and application, and belongs to the technical field of molecular detection. The invention provides a detection reagent for chilo suppressalis organophosphorus insecticide resistance, the reagent comprises a specific RPA2 primer pair designed for chilo suppressalis acetylcholin esterase ace-1 gene A314S resistance mutation site and crRNA, a double screening mechanism of RPA amplification and CRISPR-Cas12a is combined, wild type and mutant type genes can be accurately distinguished, and the detection reagent can be used for detecting the insecticide resistance of the chilo suppressalis organophosphorus insecticide resistance to the chilo suppressalis organophosphorus insecticide resistance to the chilo suppressalis organophosphorus insecticide resistance to the chilo suppressalis organophosphorus insecticide resistance to the chilo suppressalis. The rapid, sensitive and visual nucleic acid detection of the chilo suppressalis ace-1 gene A314S resistance mutation site is realized, and a visual detection means which is high in specificity, simple and convenient to operate, economical and efficient is provided for field on-site determination of chilo suppressalis resistance populations.

The invention relates to the technical field of plant genetic engineering, and particularly discloses an alfalfa salt-alkali-resistant and low-temperature composite stress-resistant gene MsNCED5 and application of encoded protein thereof in improving salt-alkali-resistant and low-temperature composite stress-resistant capability of plants. The nucleotide sequence of the gene disclosed by the invention is as shown in SEQ ID NO.1, the full length is 1833 bp, and 610 amino acids are encoded. According to the invention, an MsNCED5 overexpression vector is constructed and medicago sativa is converted, so that an MsNCED5-OE transgenic line is successfully created, and the MsNCED5 gene expression quantity of the MsNCED5-OE transgenic line is obviously increased compared with that of a wild type. Under the salt-alkali and low-temperature composite stress, the transgenic plant has stronger stress tolerance compared with the wild plant, which indicates that the overexpression of the MsNCED5 gene improves the resistance of the plant to the salt-alkali and low-temperature composite stress. The MsNCED5 gene disclosed by the invention provides an important theoretical basis for researching an alfalfa stress resistance molecular mechanism and breeding.

The invention discloses a rice salt stress resistant gene OsHAK11 coding protein and application thereof, and belongs to the field of plant genetic engineering. According to the method, a rice OsHAK11 gene (the nucleotide sequence is shown as SEQ ID NO.1) is knocked out through a CRISPR / Cas9 gene editing technology, and a mutant with significantly enhanced salt tolerance is obtained. The gene editing vector pEGCas9Pubi-B-OsHAK11 contains sgRNA of a region as shown in a target SEQ ID NO.4, a rice receptor material is transformed through agrobacterium tumefaciens mediation, and a plant with the OsHAK11 gene subjected to frame shift mutation is obtained through screening. A salt stress experiment shows that the survival rate of the mutant oshak11 is obviously higher than that of a wild type. The invention provides a new gene resource and an efficient technical means for salt-tolerant breeding of rice.

The invention discloses an application of an OsPES gene and an encoding protein thereof in regulating and controlling the tiller number of rice, and belongs to the technical field of biology. According to the application disclosed by the invention, two OsPES BRCT structural domain homozygous knockout strains Ospesc-cas9-1 and Ospesc-cas9-5, two OsPES overexpression strains OE-OsPES-3 and OE-OsPES-7 and two RNAi (Ribonucleic Acid Interference) interference strains RNAi-OsPES-1 and RNAi-OsPES-6 are respectively obtained by taking japonica rice'cloud guide 'as a background material and utilizing related technologies. Phenotypic observation and statistical analysis find that compared with a wild type, the tiller number of an overexpressed OsPES strain is increased, and the tiller number of an OsPES interference strain and the tiller number of a BRCT structural domain knockout strain are reduced. It is found for the first time that OsPES can positively regulate the tiller number of rice and has application significance in regulation of the tiller number of rice and new strain breeding.

The invention provides an RNA polymerase variant and application thereof, and relates to the technical field of biology. Wild type T7 RNA polymerase is modified to obtain the T7 RNA polymerase variant, the enzyme specific activity of the T7 RNA polymerase variant is remarkably improved compared with that of the wild type T7 RNA polymerase, the addition amount of the polymerase in the in-vitro transcription reaction process can be reduced, and the cost is saved; in addition, the mutant also effectively reduces the generation of incomplete fragment impurities, and the mRNA integrity is significantly improved to 99%. The invention further provides a method for preparing RNA through in-vitro transcription.

The invention relates to the technical field of plant stress resistance screening, in particular to a method for screening drought-resistant mutants after EMS mutagenesis of bamboo reed cluster buds, which comprises the following steps: S1, performing hydroponic seedling strengthening on wild tissue culture seedlings and EMS-treated rooting tissue culture seedlings to obtain wild bamboo reed seedlings and EMS-treated bamboo reed seedlings; s2, screening suitable concentration of PEG-6000: screening wild type bamboo reed seedlings by using a series of PEG-6000 solutions with different concentrations, comparing survival rates of the bamboo reed seedlings under different treatments, and screening suitable concentration for simulating drought stress by a PEG-6000 water culture method; s3, simulating drought stress conditions by using the screened PEG-6000 solution with proper concentration as a permeable medium, and screening the EMS treated bamboo reed seedlings to obtain drought-resistant plants. The screening method provided by the invention is not limited by seasons, has high accuracy, and is a simple, convenient and rapid method for screening drought-resistant bamboo reed plants.

The invention belongs to the technical field of plant gene engineering, and particularly relates to an OsNUC1 gene and application of a gene mutant constructed by the OsNUC1 gene to regulation and control of rice plant height, leaf length and panicle type. The regulation is to regulate the plant height and leaf growth of the rice and influence the shape of the ear part. Compared with the prior art, the OsNUC1 gene mutant has the following advantages: (1) the OsNUC1 is directionally edited by utilizing a CRISPR / Cas9 gene editing technology, an OsNUC1 gene mutant rice material is obtained, the biological phenotype of the OsNUC1 gene mutant rice material is investigated, the phenotype investigation on T1-generation seedlings of the OsNUC1 shows that the plant heights and the leaf lengths of one-week-old and two-week-old seedlings of the mutant are obviously higher than those of wild seedlings, the leaf widths have no obvious difference, and the OsNUC1 gene mutant rice material has a good application prospect; the result shows that the OsNUC1 can regulate the plant height and the leaf growth of the rice and influence the shape of the ear part. (2) it is proved that the nucleolin gene OsNUC1 is negatively regulated by a transcription factor OsARF7, and the phenotypes such as the plant height, the leaf length, the grain length and the grain width of the OsNUC1 / OsARF7 mutant are more obvious than those of the OsNUC1 mutant. The invention further proves the application of the OsNUC1 gene in regulation and control of rice plant height and leaf growth.

The invention belongs to the technical field of molecular biology and genetic engineering, and particularly relates to a passion fruit PeDREB1 gene and application thereof in regulating temperature and drought stress tolerance. By overexpressing the passion fruit PeDREB1 transcription factor in the plant, the tolerance of the transgenic plant to temperature stress (low temperature and high temperature) and drought stress can be obviously enhanced, the growth recovery capability of the transgenic arabidopsis thaliana overexpressed with PeDREB1 after low temperature stress is obviously superior to that of a wild type, and the leaf area growth rate is increased by about 0.45-2.28%; under the condition that 0.2 mol / L to 0.3 mol / L mannitol simulates drought stress, the seed germination rate is increased by 5.39 percent to 9.29 percent, and the root length growth rate is increased by 1.08 percent to 6.72 percent; the invention provides a clear gene target and a directly applicable molecular resource for plant stress resistance molecular breeding, can provide theoretical support for subsequent clarification of a regulation mechanism of plant response temperature and drought stress, and also lays a foundation for plant stress resistance genetic improvement and new germplasm creation.

The invention provides an RNA polymerase variant and application thereof, and relates to the technical field of biology, the enzyme specific activity of the variant is obviously improved compared with that of a wild type, and the addition amount of polymerase is reduced, so that the cost is saved. Meanwhile, the mutant also effectively reduces the generation of incomplete fragment impurities and improves the integrity. In addition, the invention further provides a method for generating RNA through in-vitro transcription.

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 quinoa CqARF2 gene mutant as well as an identification method and application thereof. Chenopodium quinoa seeds are mutated on a large scale by utilizing a chemical mutagenic agent EMS, a CqARF2 gene function deletion mutant arf2 is adopted as a quinoa mutant, G at the 1050th site of the CqARF2 gene of the mutant is mutated into A, and amino acid at the 350th site is mutated into a termination codon, so that the translation process of CqARF2 protein is terminated in advance. The chenopodium quinoa CqARF2 gene function deletion mutant arf2 shows remarkable agronomic character improvement, seeds are large and full, compared with a wild type, the hundred-grain weight of the mutant seeds is increased by about 40%, the seed diameter is increased by about 10%, the seed thickness is increased by about 30%, and a certain foundation is laid for breeding new high-yield and high-quality chenopodium quinoa varieties.

Provided are a gene regulating a leguminous plant plant type having tolerance to high density, and a use thereof. The nucleotide sequence of the gene is as shown in SEQ ID NO: 1, the amino acid sequence of a protein encoded thereby is as shown in SEQ ID NO: 2, and the genome sequence is as shown in SEQ ID NO: 3. Transgenic leguminous plants having the gene have a reduced branch number, a reduced plant height, a reduced node number, a shortened internode length, and / or a shortened petiole length compared to control or wild-type plants.

The invention discloses a cold response PaIBH1 protein and application of a coding gene thereof in enhancing cold stress resistance of plants. The cold response PaIBH1 protein is a protein PaIBH1 with an amino acid sequence as shown in SEQ ID NO: 2. Experiments prove that the gene for coding the protein PaIBH1, namely the gene PaIBH1, is overexpressed in wild type arabidopsis thaliana, so that the cold resistance of the arabidopsis thaliana can be improved; the improvement of the cold resistance shows that after cold treatment, the fresh weight of the overground part is increased, the wilting rate is reduced, the leaf membrane damage and electrolyte leakage are reduced, the photosynthetic ability is improved, the membrane lipid peroxidation damage is relieved, the active oxygen scavenging ability is enhanced and / or the active oxygen accumulation in the plant body is reduced. Therefore, the protein PaIBH1 can improve the cold resistance of the plants. The method has a very wide application prospect in the aspect of cold-resistant genetic improvement of crops or pasture.

Disclosed is a Taq DNA polymerase mutant. Compared to a wild-type Taq DNA polymerase, the Taq DNA polymerase mutant has a mutation at one or more of amino acid positions 116, 439, and 468, wherein the amino acid sequence of the wild-type Taq DNA polymerase is set forth in SEQ ID NO: 1. Compared to the wild-type Taq DNA polymerase, the Taq DNA polymerase mutant of the present invention has higher enzyme activity than the wild-type enzyme; both single-site mutants and multi-site mutants show improved tolerance to corn leaf supernatant, mouse serum, and / or heparin sodium, and thus can be used for direct rapid amplification of samples.