1979 results about "Wild type" patented technology

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 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 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 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 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 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 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 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.

The invention provides a method for constructing a rice epigenetic recombinant inbred line, and belongs to the technical field of molecular biology and genetic breeding. According to the method for constructing the rice epigenetic recombination inbred line, a fertile rice DDM1 gene function weakening mutant is obtained through a CRISPR / Cas9 gene editing technology and hybridized with wild type rice with the same background to obtain F1, then the F1 and the wild type rice are subjected to backcrossing to obtain BC1F1, single-plant offspring selfing with the DDM1 genotype being the wild type is selected, selfing is conducted for multiple generations through a single-grain transmission method, and the rice epigenetic recombination inbred line is obtained. According to the present invention, the strain population with stable inheritance is constructed, such that the rice epigenetic recombinant inbred line is obtained, and has characteristics of rich phenotype, genetic and epigenetic variation so as to provide important population resources for batch identification and the like of important agronomic shape functional sites of rice.

The invention provides a structure, a sequence and a construction method of a plant virus gene editing vector and application of the plant virus gene editing vector in plant gene editing. The method is characterized in that a TRV2 genome of a tobacco embrittlement virus (TRV) is used for carrying and expressing a small Cas gene AsCas12f and gRNA of the small Cas gene AsCas12f. Preferably, the AsCas12f and the gRNA of the AsCas12f are subjected to tandem expression by utilizing the TRV2 to form an expression cassette structure of the AsCas12f-tRNA-gRNA, the expression cassette substitutes a part of 2b gene sequence of the TRV2 on a TRV2 genome to form a plant gene editing vector pTRV2-Cas12f-gRNA based on the TRV virus, and long-time stable expression and gene editing can be performed after wild type nicotiana benthamiana is inoculated. The invention also provides a method for carrying out gene editing in a wild type plant by utilizing the plant virus.

The invention discloses a dihydrodaidzein reductase mutant and application of the dihydrodaidzein reductase mutant in synthesis of (3S, 4R)-tetrahydrodaidzein, and belongs to the technical field of biology. According to the mutant, wild-type dihydrodaidzein reductase DHDR is taken as a parent, single-point mutation or combined mutation is carried out, and a DHDR mutant is obtained; the parent site of the single point mutation is one of the 120th site, the 122nd site, the 170th site, the 178th site, the 214th site, the 219th site, the 220th site or the 220th site; the combined mutation adopts two modes: one mode is that a single-point mutation DHDR mutant is taken as a parent, mutation is carried out, and a combined mutation mutant I is obtained; and 2, taking the DHDR combined mutation mutant I as a parent, and performing mutation to obtain a combined mutation mutant II. The DHDR mutant disclosed by the invention is used for synthesizing (3S, 4R)-tetrahydrodaidzein by reducing dihydrodaidzein, and asymmetric reduction with high activity and high stereoselectivity can be realized.

The invention relates to the technical field of biology, in particular to application of a biological product expressed by an EsPTI15 gene to regulation and control of tiller number. The nucleotide sequence of the EsPTI15 gene is shown as SEQ ID NO: 1, it is found for the first time that the tiller number of EsPTI15 gene overexpressed siberian wildrye is remarkably increased compared with wild siberian wildrye, and the result proves that overexpression of the EsPTI15 gene can positively regulate and control the plant tiller development process. The discovery not only enriches the molecular mechanism theory of plant tillering regulation, but also verifies that the EsPTI15 gene has the application potential of increasing the tillering number of the plant, and provides key theoretical support and gene resources for subsequent technical research and development of introducing the EsPTI15 gene through gene operation, increasing the tillering number of forage grass and further realizing biomass improvement.

Methods, systems, and software for estimating circulating tumor fraction are provided. A first plurality of nucleic acid sequences for a plurality of loci in genomic DNA from a solid tumor sample is obtained. A second plurality of nucleic acid sequences for a plurality of cell-free DNA fragments obtained from a liquid biopsy sample from the same subject is obtained. One or more somatic mutations is identified in the first plurality of nucleic acid sequences. A variant allele frequency (VAF) is determined for each somatic mutation based on a frequency of the respective somatic mutation in the liquid biopsy sample and a frequency of the corresponding wild type allele in the liquid biopsy sample, thereby determining a set of VAFs. An estimate of the circulating tumor fraction for the test subject is determined based on the set of VAFs for the one or more somatic mutations.

The invention discloses a male sterility gene ZmUCE4 and application thereof in creating a male sterility line of corn, and belongs to the field of plant biotechnology breeding. According to the invention, site-directed mutagenesis of the ZmUCE4 gene in wild type corn is carried out through a CRISPR / Cas9 gene editing technology, pollen development abnormality and abortion can be caused, complete male sterility is caused, and it is proved that the ZmUCE4 gene has a regulation and control function on corn male reproductive development. Through offspring screening, a sterile line without transgenic components can be obtained, a stable maize male sterile line is created, and the method has important significance on maize male fertility control and hybrid seed production. The invention also designs a functional molecular marker aiming at the obtained uce4 male sterility mutant gene, and has important application value in maize male sterility line cultivation, sterile hybrid seed production and molecular marker-assisted selection.

The present disclosure provides guided microbial remodeling (GMR) methods for the rational improvement of plant-associated microbes to perform plant-beneficial functions. The GMR methods described herein allow for non-intergeneric genetic optimization of key regulatory networks within the microbes, which improve plant-beneficial functions over wild-type microbes but don't have the risks associated with transgenic approaches (e.g., unpredictable gene function, public and regulatory concerns, etc.). The present disclosure also provides remodeled microbes and compositions thereof. The utilization of remodeled microbes and compositions thereof will enable farmers to realize more productive and predictable crop yields without the nutrient degradation, leaching, or toxic runoff associated with traditional synthetically derived fertilizers.

The present disclosure provides codon optimized GBA1 polynucleotides encoding a GCase protein, wherein a portion of the coding sequence deviates from the wild type. The disclosure also provides expression constructs, vectors, viral particles or compositions containing the disclosed polynucleotides. In addition, methods and uses of these polynucleotides, expression constructs, vectors, viral particles or compositions are also provided, including the treatment of diseases or conditions associated with GCase deficiency.

The application belongs to the technical field of genetic engineering, and provides application of a PagLBD16.2 gene in regulating development of a poplar plant type, wherein a nucleotide sequence of the PagLBD16.2 gene is shown as SEQ ID NO. 3. The regulation is achieved by overexpressing the PagLBD16.2 gene, so that the plant height of a transgenic poplar is lower than that of a wild type 84K, the internode diameter is lower than that of the wild type 84K, and the number of internodes is more than that of the wild type 84K. The application has important significance for promoting genetic improvement and breeding of the poplar by overexpressing the poplar gene PagLBD16.2 to regulate development of the poplar plant type.

The invention discloses a carbonyl reductase mutant as well as a preparation method and application thereof, and belongs to the technical field of gene engineering and enzyme engineering. According to the invention, wild-type carbonyl reductase of Stachybotrys chlorohallonata is taken as an evolution template, and a plurality of mutants with catalytic activity, thermal stability and substrate spectrum obviously superior to those of the wild-type carbonyl reductase are obtained through an enzyme engineering technology. The carbonyl reductase mutant can catalyze various prochiral carbonyl compounds to obtain chiral alcohol with high stereoselectivity (ee is greater than 99%). The carbonyl reductase mutant provides a novel, efficient and environment-friendly technical route for industrial production of chiral alcohol products, and has very high industrial application potential.

The invention discloses application of protein derived from wheat and related biological materials thereof in dwarf wheat breeding. The TaWAK3-B gene in a wheat genome is knocked out by utilizing a gene editing technology, so that a dwarf wheat variety is cultivated, and a novel molecular breeding method is provided for improving the yield and quality of wheat. Experimental results show that compared with wild type wheat, after TaWAK3-B is knocked out, the plant height of the knocked-out line wheat is remarkably reduced, and the heading period is remarkably delayed. Therefore, the TaWAK3-B protein has the potential of regulating the plant height and the growth and development period of wheat, and can be applied to lodging-resistant breeding and plant type improvement of plants.

The invention relates to the technical field of biology, in particular to a rice salt stress regulation gene OsMTA1 and application thereof.The rice salt stress regulation gene OsMTA1 is obtained by conducting CRISPR gene editing on the OsMTA1 gene, and compared with receptor rice Zhonghua 11 (ZH11), after two independent strains with mutants knocked out of the transgenic rice OsMTA1 gene are subjected to 180mM NaCl simulated salt stress treatment, the rice salt stress regulation gene OsMTA1 is obtained. After recovery treatment, phenotype observation and survival rate statistics are carried out, it is confirmed that the osmta1 mutant has a salt-sensitive phenotype compared with wild type ZH11, and it is indicated that the OsMTA1 gene plays an important role in the process of responding to rice salt stress.

The invention relates to the field of gene engineering and molecular breeding, and discloses application of a ZmTCP5 gene in regulation and control of heat resistance of corn. According to the invention, three types of knockout mutants of the gene are obtained by using a CRISPR / Cas9 technology, and three overexpression materials are created. At normal temperature (25-28 DEG C), the material has no obvious phenotypic difference with a wild type; however, under high temperature stress (45 DEG C), the ZmTCP5 knockout mutant has enhanced heat resistance, reduced reactive oxygen species (ROS) accumulation, and significantly increased ROS scavenging enzyme activity, chlorophyll content and photochemical efficiency; on the contrary, the ZmTCP5 overexpression strain is more sensitive to heat stress, which indicates that the ZmTCP5 negatively regulates the heat resistance of the corn. The research provides a theoretical basis for analyzing a heat-resistant molecular mechanism of the corn, and also provides an important gene resource for creating a new germplasm of the heat-resistant corn.

The invention relates to the technical field of molecular biology, in particular to a qPCR (quantitative polymerase chain reaction) detection kit for detecting cis-trans mutation and a use method of the qPCR detection kit. The invention provides a primer pair which is used for detecting a first mutation site and / or a second mutation site of a target gene and comprises a universal primer F matched with the upstream of the first mutation site, a downstream primer R-BW matched with a wild type sequence of the second mutation site and a downstream primer R-BM matched with a mutation type sequence of the second mutation site, the fluorescent probe P-AW is matched with the upstream of the first mutation site, the fluorescent probe P-AM is matched with the mutant sequence of the first mutation site, and the fluorescent groups of the fluorescent probe P-AW and the fluorescent probe P-AM are different. According to the reaction system disclosed by the invention, the MGB fluorescent probe and the ARMS primer are combined, and mutation information and a cis-trans relationship can be judged according to the existence of a fluorescent signal. Compared with an existing detection method, the method for detecting the cis-trans relation through fluorescent quantitative PCR is proposed for the first time, detection can be completed within 2 h, specificity is good, the cost advantage is achieved, operation is easy and convenient, and clinical operation is convenient.

The invention provides an RNA polymerase variant, 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. In addition, the invention further provides a method for generating RNA through in-vitro transcription.

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.

The present disclosure relates to Taq DNA Polymerase mutants comprising at least three mutations which are capable of high levels of DNA amplification in the presence of several PCR inhibitors and compositions comprising the same. Also disclosed are methods of performing mutagenesis on wild-type Taq DNA polymerase to obtain the disclosed Taq DNA polymerase mutants, DNA amplification using the same, and cDNA amplification using the same.

The invention discloses application of a PdeTLP protein and a related biological material thereof in regulation and control of plant root system development. Experiments prove that compared with wild poplar, the number of adventitious roots, the number of adventitious root lateral roots, the root length and the root included angle of PdeTLP knockout poplar are obviously increased, and the plant height, the number of stem nodes and the stem diameter are also obviously increased. In addition, the xylem width, the phloem width and the xylem cell area of the poplar with the PdeTLP knocked out are obviously higher than those of the wild poplar. The PdeTLP gene participates in regulation and control of the root system number, the root system length, the root system included angle, the plant height, the stem node number, the stem diameter and the secondary microtubule tissue development of the poplar. The method not only has important significance on molecular mechanism and theoretical research of regulating and controlling the number and length of poplar root systems, plant height, number of stem nodes and stem diameter, but also has important guiding significance on directionally cultivating trees meeting actual production requirements.