413 results about "Versus gene" patented technology

The invention relates to a method for identifying gene mutation types in the field of gene analysis as well as a special chip and a kit thereof. The gene mutation detecting method comprises the following steps: taking a genome to be detected from a human tissue as a template, carrying out multiple allele special PCR amplification by a primer group that is designed aiming at special mutant sites and DNA polymerase without 3'-5' end exonclease activity, then hybridizing the obtained PCR product and an oligonucleotide probe (allele special probe) on the gene chip, and confirming mutation types of all gene sites according to the hybridizing result. The allele special probe is designed aiming at special gene types of gene mutant sites to be detected. The invention can detect gene mutations in comprehensive, systemic and high-flux ways and has light environmental pollution as well as simple and rapid operation compared with PCR-RFLP and a sequencing method.

The invention provides construction of an EP153R and EP402R gene co-expression recombinant adenovirus vector and a adenovirus packaging method, and belongs to the technical field of genetic engineering. Based on a pShuttle-CMV eukaryotic expression vector, an EP153R gene and EP402R gene overexpression adenovirus vector pAD-Shuttle-CMV-CTLA4-EP153R-EP402R is introduced with CTLA4, EP153R and EP402Rgenes at multiple cloning sites. The invention also provides recombinant adenovirus for co-expressing EP153R and EP402R genes, and the adenovirus packaging process is achieved by utilizing the constructed pAD-Shuttle-CMV-CTLA4-EP153R-EP402R vector to obtain the adenovirus capable of directly infecting animal or eukaryotic cells, thereby achieving the purpose of co-expressing EP153R and EP402R genes in eukaryotic cells, and laying a foundation for researching the adenovirus vaccine for achieving the co-expression of EP153R and EP402R genes.

The invention provides molecular markers co-separated with bruchid resistance genes VrPGIP of vigna radiate. The bruchid resistance genes VrPGIP of the vigna radiate are positioned in regions of 5.558Mb and 5.625Mb of a chromosome 5 of the vigna radiate; the molecular markers co-separated with the genes VrPGIP comprise SSR (Simple Sequence Repeat) markers Vr05-5590 and Vr05-5597; physical distances between the Vr05-5590 and the Vr05-5597 and the bruchid resistance genes VrPGIP of the vigna radiate are 1.5kb and 850bp respectively. The invention also provides application of the molecular markers in mapping of the bruchid resistance genes of the vigna radiate or genetic breeding of the vigna radiate. According to the molecular markers and the application thereof disclosed by the invention, by finely mapping the bruchid resistance genes VrPGIP of the vigna radiate, two SSR markers co-separated with the bruchid resistance genes VrPGIP of the vigna radiate are found; the development of the molecular markers provides a feasible way for assistant breeding of bruchid resistant molecules of the vigna radiate.

The invention discloses a design method of an SSR label primer. In order to overcome the defect of insufficient polymorphism of the conventional SSR label primer, the invention provides a method for designing a novel SSR label primer based on a draft sequence of a genome. The design method comprises the following steps: firstly selecting an SSR label primer with a known site as a starting SSR label primer; secondly, comparing the starting SSR label primer with the draft sequence of a chromosome to which the starting SSR label primer belongs, and finding a contig in a comparison result; thirdly, searching an SSR sequence in the contig as a finishing SSR sequence; finally, designing the SSR label primer based on the finishing SSR sequence. The invention provides 14 pairs of novel SSR label primers related to wheat stripe rust resistance, and a method for establishing a wheat genetic map with L693*L661 and L661*L693F2 single plants as a mapping population, wherein five pairs of the SSR label primers generate genetic polymorphism in a genetic test group. According to the design method, the number of the SSR label primers with known sites can be quickly increased and the polymorphism of the SSR label is increased; the genetic map can be quickly encrypted through combination with initial gene localization.

This disclosure concerns high-resolution mapping and candidate gene cloning of Rf4, a maize restorer of fertility gene that restores fertility to C-type cytoplasmic male sterility. The disclosure also relates to molecular markers that are tightly-linked to, or reside within, the Rf4 gene. In some embodiments, methods are provided whereby hybrid seeds may be produced from crosses of a male plant comprising nucleic acid molecular markers that are linked to or that reside within the Rf4 gene and a female plant carrying C-type CMS.

A method for applying adhesive tail end joints to flanking sequence separation includes following steps: 1) extracting a genome DNA (deoxyribonucleic acid) to be detected; 2) selecting isocaudarner and designing and synthesizing corresponding joints; 3) digesting the genome by the isocaudarner to form DNA segments with identical adhesive tail ends; 4) detecting digestion quality in an electrophoresis manner, quantifying the concentration of the DNA segments, connecting the joints with digestion products of the genome, and forming each DNA segment with 'the joint, the digestion segment and the other joint' sequentially; 5) purifying the DNA segments with 'the joints, the digestion segments and the other joints' sequentially, and performing PCR (polymerase chain reaction); 6) recycling the segments obtained from the step 3) and sequencing; 7) analyzing a sequencing result and obtaining an unknown flanking sequence of a known sequence; and 8) checking the unknown flanking sequence. The method for flanking sequence separation is high in connection efficiency and generality and low in cost.

The invention relates to a high-oleic-acid-content molecular marker, a back cross breeding method for carrying out assistant selection by utilizing the molecular marker, and an application of the back cross breeding method. Sequences of variant specific primers of the molecular marker are as follows: a forward primer MITE-INS-F, and a reverse primer MITE-INS-R; the sequences of non-variant specific primers are as follows: a forward primer WILD-F, and a reverse primer WILD-R. The back cross breeding method utilizing the molecular maker comprises the following steps: carrying out hybridization on a peanut parent of a gene type AABB and a parent of a gene type aabb, then carrying out cross breeding; amplifying an FAD2A gene segment by utilizing one pair of primers in a genome, and selecting a single plant containing a genotype Aa; carrying out PCR (polymerase chain reaction) detection by utilizing a primer, at a site ahFAD2B, of a peanut as a marker, and selecting a single plant containing a genotype Bb; carrying out back crossing on the single plant which is dominant in two markers, namely the single plant containing the genotype AaBb, and the parent AABB until a high-oleic-acid-content single plant which is of a genotype aabb and is consistent with a recurrent parent in other economical characters is selected from later generations, and then carrying out self-crossing and amplifying propagation. The high-oleic-acid-content molecular marker has the advantages of being simple in method, low in cost and stable in result; when the high-oleic-acid-content molecular marker is used for carrying out the back cross breeding, the accuracy of character selection can be obviously improved, the breeding cost is reduced, and the breeding efficiency is improved.

The invention belongs to the field of animal gene engineering, and in particular relates to a synthetic fusion gene for expressing mycoplasma hyopneumoniae and application. The fusion gene is characterized in that a mycoplasma hyopneumoniae P97R1 gene is connected in series with a P36 gene through a Linker, the termination codon of the P36 gene is deleted, the P46 gene with signal peptide removed is fused at the C end of the P36 gene through Linker, and then the fusion gene P97R1-P36-P46 is obtained, wherein the nucleotide sequence of the fusion gene is shown as SEQ ID NO: 1. The fusion gene is included in a prokaryotic expression plasmid, and escherichia coli BL21/pET30a-P97R1-Linker-P36-Linker-P46 containing the plasmid is collected in CCTCC with the collection number CCTCC NO: M2014269. The invention further discloses immune efficacy evaluation of the fusion gene and application of the fusion gene in a novel vaccine.

The invention discloses a method for detecting Xanthomonas oryzae pv.oryzae, Xoo, X.oryzae pv. Oryzicola, Xoox, X.axonopodis pv. Citri, Xac, and X.campestris pv.campestris, Xcc. The method comprises the following steps: carrying out PCR (polymerase chain reaction) amplification on the DNA template of a sample to be detected by using two pairs of primers at the same time; then, hybridizing the PCR products with the specific probe arranged on a gene chip; and judging whether the sample to be detected is with Xanthomonas oryzae pv.oryzae, Xoo, X.oryzae pv. Oryzicola, Xoox, X.axonopodis pv. Citri, Xac, and X.campestris pv.campestris, Xcc according to the fact whether the hybridization result is positive or not. Therefore, the method of the invention is capable of detecting four bacteria by one time with high detection speed and has the advantages of direct-viewing and clear detection result and easy judgment.

The invention relates to a pSGS vector and application of the vector to development of a high-expression cell strain. The construction of the vector comprises the following steps: based on a neomycin resistance screening gene NEO, through combined strategy of weakening screening gene expression and reinforcing target gene expression, constructing an expression vector pSNEO, and in view of the amplification function of a GS system, through designing a primer, replacing the hairpin structure sequence and the NEO gene of the vector pSNEO with a hairpin structure sequence and a GS gene to obtain a universal vector pSGS for screening a high-expression and stable-transfection cell strain. Through the pSGS vector constructured by the invention, the construction of a stable and high-expression cell strain of the target gene can be finished conveniently and quickly, and a new tool is provided for preparation of macromolecular recombinant protein.

The invention relates to a gene TaBAS1-2B related to the chlorophyll content of a leaf and the grain weight of wheat and an Indel marker of the gene. According to the gene, the gene TaBAS1-2B related to the chlorophyll content of the leaf and the grain weight of the wheat is separated and cloned from wheat for the first time, the Indel marker with polymorphism is developed between two varieties of Jing 411 and Hongmangchun 21 according to a cloned TaBAS1-2B gene sequence, the effect on phenotype is analyzed in 194 natural wheat varieties, and finally, a functional marker which is separated together with the gene TaBAS1-2B and is closely related to the chlorophyll content of the leaf and the grain weight of the wheat is developed. The Indel marker can be used for respectively explaining 8.2% and 4.4% of phenotypic variation of the chlorophyll content of a flag leaf and the thousand seed weight after wheat anthesis, wherein a strip of which the molecular weight is 494bp (Seq ID No.3) has a synergistic effect on the increase of the grain weight, and a strip of which the molecular weight is 493bp (Seq ID No.4) has a synergistic effect on the reduction of the grain weight. The development of the molecular marker provides a practical way for assistant breeding of high-yield wheat molecules.

The invention belongs to the technical field of biology, and relates to wild type short-chain dehydrogenase LfSDR1 and a gene of mutant enzyme of the short-chain dehydrogenase, construction of a recombinant expression vector containing the gene, preparation of recombinant expression transformant, recombinase and a preparation method of the recombinase. Through mutation of the wild type short-chaindehydrogenase LfSDR1, the mutant enzyme having two stereoselectivities (R or S stereoselectivities) is obtained, and catalytic asymmetrical reduction on prochiral ketone is carried out by utilizing the mutant enzyme, such as asymmetrical reduction of catalytic 1-phenyl ethanone compounds. Compared with the wild type short-chain dehydrogenase, the mutant disclosed by the invention has the advantages that the stereoselectivities is improved or reversed, a substrate is catalyzed so as to obtain chiral alcohol having two types (R or S type), such as optical pure (R)- or (S)-2-chloro-1-phenylethanol; moreover, the mutant has a good application value in the field of preparation of chiral alcohol.

The invention discloses an Escherichia coli for producing riboflavin and constructing method and use of Escherichia coli. The preparation method comprises the following steps: designing ribosome bind sites RBS1, RBS2, RBS3, RBS4 and RBS5, respectively connecting the ribosome bind sites with genes ribA, ribB, ribD, ribE and ribC into fragments, splicing the fragments in order and enzyme-cutting and linking to a plasmid pTrc99a to obtain plasmid p20C-EC10, transferring into the Escherichia coli to obtain RF01S strain, transforming related gene of glycolytic pathway and ED pathway of the RF01S strain so as to improve the output of the riboflavin, and transforming the consumption pathway of the riboflavin to decrease the metabolism velocity of the riboflavin so as to further improve the riboflavin accumulation, inserting a Ptrc strong promoter into genome of the strain to obtain the Escherichia coli for producing the riboflavin. The constructed Escherichia coli strain has clear genetic background, and the glucose can be used as substrate to produce the riboflavin, and the shake-flask fermenting result is more than 1g/L, and a foundation is provided for improving the riboflavin output and yield.

A method of site-specific modification of an endogenous target DNA of a eukaryotic cell is provided. The method includes contacting the endogenous target DNA having an intended modification site with (i) a gene editing system configured to introduce a double strand break in the endogenous target DNA at or near the intended modification site, and (ii) a donor DNA repair template comprising a plurality of tandem repeat sequences. In the method, each of the plurality of tandem repeat sequences comprises an exogenous donor DNA sequence flanked by a donor 5′ flanking sequence and a donor 3′ flanking sequence. The donor 5′ flanking sequence and the donor 3′ flanking sequence are homologous to a continuous DNA sequence on either side of the intended modification site in the endogenous target DNA.

The invention discloses a method for screening and identifying spinocerebellar ataxia type 3 (SCA3)/Machado-Joseph disease (MJD) molecular marker MicroRNAs (miRNAs) capable of regulating and controlling the expression of an ATXN3 gene. The method comprises the following steps: obtaining differential expression profile of miRNAs by using miRNA microarray chip technique; further screening and verifying by using a bioinformatics method and a qRT-PCR technique to obtain the miRNAs expressed by regulated ATXN3 gene; determining the target action relation and mechanism of action of the miRNAs and ATXN3 by using qRT-PCR technique and Westernblot; determining target action relation and specific binding site of the miR-25 and ATXN3 gene by using luciferase reporter gene detection technique so as to provide basis for researching and developing diagnostic kit related to SCA3/MJD disease.

The invention discloses a cationic silk fibroin/gene compound, and a preparation method and application thereof and belongs to the technical field of polymeric biomaterials. The cationic silk fibroin/gene compound, and the preparation method and application thereof are characterized in that a cationic silk gene transmission carrier which has cell targeting and cell nucleus positioning functions and can be biodegraded is established and is obtained through the following steps: protamine sulfate is mediated by a water-soluble 2-imino thiacyclopentane hydrochloride to be in coupled with and react with the silk fibroin activated by sulfo-succunyl imino group-4-[N-maleimide methyl]-cyclohexane-1-carboxylic ester; the combination and a gene matter form the cationic silk fibroin/ gene compound through electrostatic interaction. According to the invention, the cationic silk fibroin/gene compound has good biocompatibility and degradability, is controllable in surface charge density, can be effectively compressed and protect DNA, is relatively high in transfection efficiency, and has the particular cell targeting and cell nucleus positioning functions.