109 results about "Alpha Gene" patented technology

The invention belongs to the technical field of analysis of gene expression, and discloses application of a CmEF1 alpha gene and a CmRAN gene used as reference genes in analysis of genetic expression of Cucumis melo L. fruits. The nucleotide sequence of the CmEF1 alpha gene is shown in SEQ ID No:1; the nucleotide sequence of the CmRAN gene is shown in SEQ ID NO:2. The invention further discloses a method for analyzing genetic expression of Cucumis melo L. fruits through adopting real-time fluorescence quantification PCR, and the method includes the step that the geometric mean of the gene combination of the CmEF1 alpha gene and the CmRAN gene is used as the reference genes in analysis of genetic expression of Cucumis melo L. fruits. According to the invention, the CmEF1 alpha gene and the CmRAN gene can be stably expressed in Cucumis melo L. fruits adopting different fruit bearing manners and at different developmental stages, so as to serve as a reliable reference gene combination in analysis of genetic expression during the development process of Cucumis melo L. fruits adopting the real-time fluorescence quantification PCR technology for detection.

By using an antisense nucleotide, a ribozyme, a maxizyme, or an RNAi constructed based on the nucleotide sequence of exon 1 beta of the PDGF receptor alpha gene, which is expressed in specific cancer cells, or a polypeptide containing a portion thereof, translation of an mRNA transcribed from exon 1 beta of the PDGF receptor alpha gene is suppressed. An agent for suppressing expression containing as an active ingredient a substance for inhibiting expression, such as an antisense nucleotide, a ribozyme, a maxizyme, or an RNAi, is effective as a therapeutic agent for cancer.

The invention provides a genetic marker relevant to the high altitude hypoxia adaptability of Tibetan sheep. The genetic marker is positioned on a PPAR (peroxisome proliferator activated receptor) alpha gene, and a concrete nucleotide sequence is as shown in a sequence table SEQ ID No. 1; in the nucleotide sequence of the sequence table SEQ ID No. 1, when an allelic gene is A, the high altitude hypoxia adaptability of the Tibetan sheep is poor; when the allelic gene is B, the high altitude hypoxia adaptability of the Tibetan sheep is good. The Tibetan sheep are taken as research objects, a PCR(Polymerase Chain Reaction)-SSCP (single-stranded conformational polymorphism) method is adopted to combine with DNA (deoxyribonucleic acid) sequencing technology so as to research the genetic variation characteristics of PPAR alpha genes of 2 sheep groups, and in the high altitude hypoxia adaptability, the molecular markers of the Tibetan sheep are initially screened out to provide foundation for subsequent relativity analyses.

The invention discloses a method for improving antiviral activity of a recombinant swine interferon-alpha fusion protein. The nucleotide sequence of the fusion protein is of SEQ ID NO.1 as shown in the specification, and an amino acid sequence encoded by using the fusion protein is of SEQ ID NO.2 as shown in the specification. By adopting the method, a swine interferon-alpha gene and a swine antibody CH3 fragment are subjected to fusion expression, the stability of the protein is improved, the activity time is prolonged, and efficient preparation is achieved through a prokaryotic expression system, and tests show that the half-life period of the recombinant swine interferon-alpha expressed by using the protein is remarkably prolonged when being compared with that of natural swine interferon-alpha, and the bioactivity of the recombinant swine interferon-alpha is also prior to that of a natural product.

The present invention discloses the preparation process of thynosin-alpha with acetylation modified N-end by using recombinant colibacillus. The preparation process includes obtaining thynosin-alpha gene, constructing recombinant expression vector containing thynosin-alpha, transforming prokaryotic cell, culturing prokaryotic cell and expressing thynosin-alpha, and separating and purifying thynosin-alpha with acetylation modified N-end. The present invention also discloses the process of utilizing recombinant colibacillus in preparing thynosin-alpha antigen with acetylation modified N-end and then cutting and separating to prepare thynosin-alpha-1 antigen with acetylation modified N-end, thynosin-alpha-2 antigen with acetylation modified N-end and similar matter. The thynosin-alpha with acetylation modified N-end has the functions of regulating immunity, cell proliferation, etc. and has wide application foreground in antivirus and antitumor.

The invention discloses an IKK alpha gene RNA Interfere mouse model mediated by lentiviral vector and usage. The experiment constructs the lentiviral vector aiming at IKK alpha gene RNAi (knockdown), the vector is inducted into perivitelline space of fertilized eggs of a mouse to obtain transgenic mouse model which carries the vector, and the average positive rate of founder mice is 15% via the verification of a PCR. Then detection of a reverse transcription PCR and a method of real-time quantitative PCR shows that the expression level of peripheral blood cells IKK alpha mRNA of the transgenic mice is obviously decreased. The biological function of IKK alpha gene and the relation between the abnormal expression and tumor can be researched in vivo by employing the model, thereby finding anti-tumor drugs.

The invention relates to a PPAR (Peroxisome Proliferator-Activated Receptor) alpha gene and application thereof as goose fat traits genetic markers. Genetic markers related to abdomen fat weight and abdomen fat ratio in the PPAR alpha gene are measured; the existence of polymorphism in the PPAR alpha gene in a nucleic acid sample obtained from a goose is measured, wherein the polymorphism is related to the abdomen fat weight and the abdomen fat ratio; and the fat traits selection breeding can be carried out on the goose containing the PPAR alpha gene in a genome DNA according to the single nucleotide polymorphism of the gene, and an advantageous genotype for reducing the abdomen fat weight and the abdomen fat ratio is obtained by the screening. The invention has extremely important application value and economic benefit.

The invention relates to a plant expression vector and a construction method thereof, and a method for producing chicken alpha interferon by utilizing crowtoe as a bioreactor, belonging to the technical field of biology. The plane expression vector pSFRLH is shown as the figure 1 in the specification. In the invention, main stem sections, rachis stem sections and tender leaves of crowtoe are used as explants, and a constructed plane expression vector chicken alpha interferon gene ChIFN-alpha is subjected to genetic transformation and regeneration and then subjected to PCR (Polymerase Chain Reaction), RT-PCR (Reverse Transcription-Polymerase Chain Reaction), Southern blot, Western blot and ELISA (Enzyme-Linked Immuno Sorbent Assay). Proved by a detection result, the ChIFN-alpha gene is integrated into a crowtoe chromosome genome and correctly transcribed, translated and expressed, and recombinant protein can be neutralized by a ChIFN-alpha antibody. The invention lays the foundation for producing animal medical protein by utilizing crowtoe pasture grass as a bioreactor and provides a new thinking for the control of animal epidemic diseases.

The invention relates to a method for preparing polypeptide vaccine agent used in foot-and-mouth disease gene project, wherein said agent is recombined particle that prepared by colonizing pig interferon-alpha gene via molecule biological technique into eucaryon expression carrier; the said agent is recombined particle that prepared by colonizing pig is one of interferon-alpha gene group; the eucaryon expression carrier is any DNA expression carrier with eucaryon starter, protocaryon copier, and protocaryon resistant gene; said inventive particles will transfer into bacillus coli to expand, via alkali crack method to obtain many particles, to be diluted by PBS buffer liquid into some density, to prepare inventive polypeptide vaccine agent. Animal test has proved that said agent is safe coupled with O-tyep vaccine of foot-and-mouth disease gene project, while the protection ration can reach 100% by one injection.

The invention discloses a probe and a gene chip for thalassemia detection, a preparation method and applications. The preparation method of the probe for the thalassemia detection includes the following steps: constructing a capture interval library based on different thalassemia genes, modified genes and mutations; and designing a capture probe according to the capture interval library, wherein the capture interval library includes all globin genes and important regulatory region full-length fragments of an alpha gene cluster and a beta gene cluster, recorded breakpoint mutation sites, SNP sites selected from deletion mutations, and SNP sites on thalassemia modified genes. The preparation method of the invention is to design the probe aiming at all gene clusters and related genes of thalassemia, and so the probe can cover all thalassemia gene mutations. The preparation method is combined with Tn5 library construction and high-throughput sequencing, and so all the mutations can be covered, and the method has the advantages of simple operation, high throughput and low cost, and is suitable for large-scale population diagnosis and screening of thalassemia and research and predictionof new unknown gene mutations.

The invention discloses an engineering bacterium for efficiently expressing recombinant ChIFN-alpha. The engineering bacterium is obtained by the following steps of S1, performing codon optimization synthesis on the ChIFN-alpha gene sequence published in Genebank according to the preference of the colibacillus codon; S2, designing a specific primer according to the optimized ChIFN-alpha gene; S3,performing PCR amplification on the ChIFN-alpha gene; cloning the PCR products onto prokaryotic expression vectors pET-41a through EcoRI and HindIII double enzyme digestion; S4, converting pET-41a-ChIFN-alpha recombinant plasmids with the correct sequencing into BL21(DE3) competent cells; S5, performing recombinant ChIFN-alphaprokaryotic expression and antiviral activity determination. The engineering bacterium disclosed by the invention can realize the efficient expression of the ChIFN-alpha; the antiviral activity of the obtained ChIFN-alpha can reach 1.71*10<7> UI / mg.

The invention discloses an application of a DGK alpha (diacylglycerol kinase alpha) gene-chitosan nanoparticle as a unique active component in preparing a medicine for treating allergic asthma as well as a preparation method of the DGK alpha gene-chitosan nanoparticle. The invention, aiming at shortcomings that naked DGK alpha plasmid is easy to be damaged by nuclease, acid, alkali and the like in the body, prepares the DGK alpha gene-chitosan nanoparticle through chitosan, so as to effectively protect the DGK alpha plasmid, enhance the activity of the DGK alpha plasmid in the body and promote the DGK alpha gene to develop better function in the body. The DGK alpha, based on a function of induction of T cell anergy, can induce immune tolerance. The DGK alpha gene-chitosan nanoparticle prepared by the invention can inhibit egg albumin-induced airway inflammation of asthmatic mice, reduce serum egg albumin specific IgE level, and is expected to be used for treating asthma and other allergic diseases.

The invention discloses a chicken muscle fiber-type property-related gene molecule marker and a use thereof. A 646th nucleotide G-to-A mutational site of a PGC-1 alpha gene exon is used as a molecule marker, the molecule marker has a nucleotide sequence shown in the formula of SEQ ID NO. 1, and at 97th bp of the nucleotide sequence shown in the formula of SEQ ID NO. 1, G97-A97 base mutation is produced. The molecule marker provides a fast, simple and low cost gene analysis method for breeding of chicken with chicken muscle fiber-type properties, is conducive to screening of chicken with high slow muscle fiber content and good meat quality and can guide molecule marker-assistant breeding of chicken with chicken muscle fiber-type properties.

The present invention relates to a method for predicting the responsiveness of a patient to a treatment with a TNF-alpha blocking agent, said method comprising determining the presence or absence of a guanine at position −238, a guanine at position −308, and a cytosine at position −857 of the TNF-alpha gene of said patient, wherein the simultaneous presence of a guanine at position −238, a guanine at position −308, and a cytosine at position −857 of the TNF-alpha gene in both copies of said TNF-alpha gene of said patient is indicative of a lessened likelihood of responsiveness of said patient to a treatment with a TNF-alpha blocking agent with respect to standard responsiveness.

The invention discloses a PML-RAR alpha fusion gene detection kit and a detection method thereof. The kit comprises a first group of probes for detecting an upstream gene sequence of an L-type breakage hot spot of a PML gene and a second group of probes for detecting all gene sequences of a RAR alpha gene. Each one of probes is labeled a dye. The dyes labeling the two groups of probes are in different colors. The two groups of probes are amplification products obtained by amplification based on a human epigenome DNA as a template and amplification primers. The probes have strong specificity and low background noise and realizes the optimal balance between the detection specificity and hybridization time. The PML-RAR alpha fusion gene detection kit guarantees result specificity and sensitivity, shortens hybridization time, has detection time of 7-8h and improves detection efficiency.

The invention belongs to the field of molecular biology, and particularly relates to a luffa cylindrica.L reference gene EF-1 alpha and a primer and application thereof. The nucleotide sequence of theluffa cylindrica.L reference gene EF-1 alpha is shown as SEQID No.1. A luffa cylindrica.L real-time fluorescent quantitation PCR primer designed with the gene sequence is shown as SEQID No.2-3. BestKeeper, GeNorm, NormFinder and RefFinder analysis software and a Delta Ct method are adopted for evaluating the stability of the EF-1 alpha gene, and the reference gene EF-1 alpha is the most stable under high temperature and low temperature and ABA coercion condition. The designed real-time fluorescent quantitation PCR primer is high in specificity, and has high stability, reliability and repeatability, a strong support is provided for precise quantitation of relevant functional groups under adversity stress of temperature stress, ABA coercion and the like of luffa cylindrica.L, and research stability, repeatability and reliability are improved.

The invention belongs to the field of molecular biology and in particular relates to a winter squash EF1-alpha gene and application thereof. The winter squash EF1-alpha gene has a nucleotide sequenceas shown in SEQ ID No.1. Winter squash real-time fluorescent quantitative PCR (Polymerase Chain Reaction) primers designed according to the gene sequence are shown in SEQ ID No.2 and SEQ ID No.3. Thereal-time fluorescent quantitative PCR primers designed by the invention are good in specificity and very high in stability, reliability and repeatability. Tests show that the winter squash EF1-alphagene can be stably expressed in different tissues at different phases under different abiotic stress conditions, and is suitable for being used as a reference gene in winter squash gene expression research.

The invention provides to a method for constructing a recombinant rhabdovirus expression vector for EGFP (enhanced green fluorescent protein) gene, relating to a method for constructing the recombinant rhabdovirus expression vector for the EGFP gene. The invention aims to solve the problems of low transduction efficiency, large distance in expression efficiency from requirement of industrial production, short expression time and the like of the traditional recombinant rhabdovirus expression vector. The method comprises the following steps: firstly, amplifying EF1 alpha gene through PCR (polymerase chain reaction); secondly, constructing a plasmid pT-EF1 alpha; thirdly, constructing a plasmid pWK; fourthly, amplifying L-ITR (L-inverted terminal repeat) gene through PCR, and constructing pWK-L-ITR; fifthly, amplifying R-ITR gene through PCR, and constructing pWK-ITR; sixthly, amplifying the EGFP gene through PCR, and constructing the recombinant rhabdovirus expression vector pWK-ITR-EGFP for the EGFP gene. In the invention, through the addition of elements of VSV (vesicular stomatitis virus)-GED, WPRE (Woodchuck Posttranscriptional Regulatory Element) and ITR, the expression efficiency of exogenous genes is enhanced, the transduction efficiency of virus is improved, and the expression time is prolonged.

The invention discloses a method for removing USEPA PAHs in agricultural soil by using an aboriginal PAHs degradation bacterial community. The method comprises the steps of domesticizing and enriching autochthonous bacteria in soil by using PAHs as the only carbon source, monitoring the abundance of polycyclic aromatic hydrocarbon dioxygenase alpha subunit (RHD alpha) and the bacterial community changes in a culture solution, sorting appropriate bacterial community, performing extended culture, and performing inoculating in soil to reduce PADs content in the soil. The aboriginal PAHs degradation bacterial community has high-content RHD alpha genes. According to the method, USEPA PAHS in polluted farmland can be effectively removed by increasing the abundance of the RHD alpha gene in nidA and nahAc in agricultural soil, and the method has the advantages of being high in efficiency and environment-friendly.

The invention relates to a design, amplification and sequencing method of four pairs of Floccularia luteovirens nuclear gene primers. The method comprises steps as follows: (1), universal primers matched with EF1-alpha, RPB1, RPB2 and Mcm7 genes are adopted to amplify Floccularia luteovirens, and amplified fragments of the EF1-alpha, RPB1, RPB2 and Mcm7 genes are obtained respectively; (2), the amplified fragments of the EF1-alpha, RPB1, RPB2 and Mcm7 genes are subjected to gel detection, target bands are cut and subjected to sequencing after purification and recovering by a SanPrep column type DNA gel recovery kit, and 521 bp EF1-alpha gene fragments, 751 bp RPB1 gene fragments, 782 of RPB2 gene fragments and 688 bp Mcm7 gene fragments are obtained; (3), 30 bp sequences of two ends of each of the EF1-alpha, RPB1, RPB2 and Mcm7 gene fragments obtained in the step (2) are removed, the primers are designed online, and a pair of primers with the highest score is selected; (4), the specificity of the primers in the step (3) is verified through amplification and sequencing of Floccularia luteovirens strains with different geographical distribution sources. The method is simple and easy, and the primers have excellent amplification performance.

The invention relates to a 100-liter close type photo-bioreactor and a processing technique for the mass cultivation and promotion of the growth and the gene expression of trans-TNF-Alfa anabaena by using the photo-bioreactor. The reactor comprises three built-in light transmitting glass tube, an adjustable illumination facility, a sterile air blowing facility, a high temperature sterilizing system, and CO2 distribution facility. A culture medium is prepared, and regulated and controlled artificially in the reactor so as to promote the growth and the gene expression of the trans-TNF-Alfa anabaena. The production method is suitable for large scale production of transgene blue algae, and has the advantages of high stability, strong controllability and maneuverability, small outcome difference between batches, etc.

The invention discloses a reagent kit for detecting personal habitual abortion genetic susceptibility and an application method thereof. The reagent kit comprises six specific primer couples of single nucleotide polymorphic loci genetic type and specific fluorescence probe couples for detecting the associated antigen 4-gene (CTLA4) of cytotoxicity T lymphocyte, tumor necrosis factor-alpha gene (TNF-alpha), interleukin-6 gene (IL-6), and 5, 10-methylene tetrahydrofolate (MTHFR), and general components for fluorescence quantitative PCR detection. The reagent kit can be used to evaluate personal habitual abortion genetic susceptibility.

The invention discloses a PML-RAR alpha fusion gene detection kit and a detection method thereof. The kit comprises a first group of probes for detecting upstream and downstream sequences of a PML gene and a second group of probes for detecting upstream and downstream sequences of a RAR alpha gene. Each one of the probes is labeled with a dye. The dyes labeling the two groups of probes are in different colors. The two groups of probes are amplification products obtained by amplification based on a human genomic DNA as a template and amplification primers. The length of the probes is designed according to a lot of experiments. Through experiment result comparison and statistic analysis, the optimal length is obtained and the optimal balance between the detection specificity and hybridization time is realized. The PML-RAR alpha fusion gene detection kit guarantees result specificity and sensitivity, shortens hybridization time, has detection time of 7-8h and improves detection efficiency.

The invention discloses a mutant type TNF-alpha gene the base composition of which is shown as the SEQ ID NO 7, and discloses a method of increasing the expression quantity of the antigen TNF-alpha gene. The method includes steps of: amplifying by adopting a human secreting type TNF-alpha gene cDNA clone as a template and by adopting the SEQ ID NO 1 and the SEQ ID NO 2 as primers, so as to obtain an amplification product T1; amplifying by adopting the SEQ ID NO 3 and the SEQ ID NO 4 as primers, so as to obtain an amplification product T2; performing overlapping PCR amplification by adopting the amplification product T1 and the amplification product T2 as templates and adopting the SEQ ID NO 1 and the SEQ ID NO 4 as primers, so as to obtain the mutant type TNF-alpha gene; subjecting the mutant type TNF-alpha gene to double digestion; inserting into a pET28a expression vector; converting escherichia coli; and performing induced culture to obtain TNF-alpha protein. Two pairs of escherichia coli rare codons close with each other in the TNF-alpha gene mutant into escherichia coli optimal codons. The expression quantity of the TNF-alpha gene after the codon optimization is increased by at least 1.5 times.

The invention relates to a kit for Chinese pediatric asthma susceptibility gene SNP (single nucleotide polymorphism) genotyping and an application method of the kit for Chinese pediatric asthma susceptibility gene SNP genotyping, in particular to a kit for simultaneous qualitative genotyping of nine SNP loci of eight Chinese pediatric asthma susceptibility genes and an application method of the kit for simultaneous qualitative genotyping of the nine SNP loci of the eight Chinese pediatric asthma susceptibility gene. Genotypes of the nine SNP loci are obtained simultaneously by taking an rs1042713 locus of an ADRB2 gene, an rs1805010 locus of an IL-4R alpha gene, an rs20541 locus of an IL-13 gene, an rs1800925 locus of the IL-13 gene, an rs2495636 locus of an IL-13R alpha gene, an rs2243250 locus of an IL-4 gene, an rs1800629 locus of a TNF-alpha gene, an rs569108 locus of an MS4A2 gene and an rs528557 locus of an ADAM33 gene as detection objects, designing amplification primers and extension primers respectively aiming at mutation of the nine SNP loci, performing multiplex PCR (polymerase chain reaction) amplification and marker extension aiming at the nine SNP loci, and performing capillary electrophoresis. The kit for SNP genotyping is convenient to use, simple to operate, low in cost, high in flux and direct and reliable in detection results and is applicable to large-scale screening of Chinese pediatric asthma susceptibility gene SNP genotyping.

The invention discloses a plasmid vector, a building method thereof and an application. PCAMBIA1300 is modified to form the plasmid vector, botryosphaeria dothidea histone H3 gene promoters, botryosphaeria dothidea transcription elongation factor alpha gene promoters and resistance genes are inserted into pCAMBIA1300 multiple cloning sites, and the resistance genes are positioned at downstream positions of the botryosphaeria dothidea transcription elongation factor alpha gene promoters. According to the plasmid vector, segments among botryosphaeria dothidea histone H3 genes and the transcription elongation factor alpha gene promoters are selected, and genes connected at the back can be efficiently started, so that conversion efficiency of the plasmid vector is greatly improved. According to the building method, the botryosphaeria dothidea is efficiently converted by the aid of exogenous genes mediated by agrobacterium tumefaciens, and a feasible scheme is provided for research and development of aspects such as genetic modification of the botryosphaeria dothidea, development of interested genes, drug target genes separating pathogenic fungi and development of gene functions of the botryosphaeria dothidea.

The invention discloses a kit for detecting whether relative SNP sites on habitual abortion pathogenic genes mutate or not. The kit provided by the invention mainly comprises the following components of: (1) specific primer pairs and specific fluorescent probe pairs for detecting the genotype of polymorphism of No.rs769214 SNP site on CTLA4 gene, No. rs1800629 and No. rs1799724 SNP sites on TNF-alpha gene, No. rs1800796 SNP site on IL-6 gene, No. rs1801133 and No. rs1801131 SNP sites on MTHFR gene; (2) a routine fluorescence quantitative PCR reaction reagent. The kit provided by the invention is used for detecting the mutant of habitual abortion pathogenic genes so as to rapidly and conveniently check out pathogenic gene carriers and patients, and can be applied in antenatal diagnosis and timely treatment so as to reduce the incidence of habitual abortion and improve pre-natal and post-natal care.

The invention discloses a method for detecting bastard halibut LITAF gene expression by applying reverse transcription-polymerase chain reaction (RT-PCR). The method lays a foundation for studying bastard halibut LITAF gene expression regulation mechanism and immunology function. TNF-alpha plays an important role in the process of immune response and foreign pathogenic bacteria killing, and the LITAF is a transcription factor of TNF-alpha gene and directly controls the expression of the TNF-alpha gene. Therefore, the expression abundance of the LITAF gene reflects the activity of a bastard halibut immune system to a certain extent and can be used as an immune monitoring index and immune agent quality evaluation index in prevention and treatment of bastard halibut diseases. By detecting the variation of the bastard halibut LITAF gene expression quantity, whether the bastard halibut is infected with a disease can be judged in advance and a measure of prevention and treatment can be taken timely so as to avoid unavoidable loss resulting from serious situation; and besides, the advantages and disadvantages of a fish immune agent can be evaluated so as to judge which immune agent has better effect during prevention and treatment of flounder diseases. The invention provides a technology platform for relative quantitative analysis of LITAF gene at the mRNA level.