41 results about "GAPDH Gene" patented technology

The present invention relates to a streptococcus adhesion antigen GAPDH and a preparation method thereof. The GAPDH is an rGAPDH, which has high conservative property and sequence homology, consists of 319 amino acid residues, and has a molecular weight of 32.4kDa. The rGAPDH is encoded by a gapdh gene; and a forward primer and a reverse primer of the gapdh gene respectively have one restriction enzyme cutting site at 5' and 3'. The preparation method of the GAPDH comprises the following steps of: PCR amplification, connection to a carrier, transformation and induction, and purification. The rGAPDH and an SEZ (Streptococcus zooepidemicus) rehabilitation pig serum show good immune responses; the rGAPDH can protect SEZ infected mice with limitation; and the mice immunized by the rGAPDH show high level of antibody titer in serum. An anti-GAPDH antibody can induce high level of bactericidal capability; a gapdh gene has a transcription level in SEZ infected mice much higher than that of culture in vitro; and the rGAPDH can significantly weaken cell adhesion ability of the SEZ, and greatly reduce cell infection ability of the SEZ.

The invention discloses a screening method of reference genes in a skin tissue of sheep, which relates to the technical field of biological engineering. The screening method comprises the steps that: a method of the reference genes is confirmed; a given amount of RNA (ribonucleic acid) extract is taken and diluted by 2-10 times by ddH2O (double-distilled water); a 50 nanograms / microlitre RNA sample is taken to serve as a template, and a cDNA (complementary deoxyribonucleic acid) synthetic reagent kit is adopted for conducting inverse transcription to synthesize cDNA; the cDNA synthesized through the inverse transcription serves as a template, and six reference genes serve as primers; and finally, pairing variation analysis of normalized factors Vn / n+1 of the six reference genes are calculated based on a geNorm program, so as to decide the amount of the optimum reference genes. The screening method solves the problem that no screening method for GAPDH (reduced glyceraldehyde-phosphate dehydrogenase) genes in the skin tissue of a Chinese merino (Xinjiang type) serving as the reference genes is not available at present. Through the combination of the specificity of the SYBR Green I with double chain DNA to generate fluorescence, a real-time fluorescent quantitative PCR (polymerase chain reaction) detection method of the SYBR Green I for the GAPDH genes in the skin tissue of the Chinese merino (Sinkiang type) is established.

The invention discloses a molecular detection method of panonychus citri Mc Gregor response abamectin stress protein encoding gene. The molecular detection method comprises the following steps: firstly, extracting total RNA (Ribonucleic Acid) of panonychus citri Mc Gregor treated by abamectin, and then transcribing the RNA into cDNA (complementary Deoxyribonucleic Acid); secondly, carrying out real-time quantitative detection on RPSA, EEF2, ATPsyn-alpha, FAH, V-ATPsyn-B, CALR, sHsp, CarE-B, Actin, CRY, Fer, Vg, ATPsyn-D, TPX, TCP-1-eta and GAPDH genes by using a qPCR (quantitative Polymerase Chain Reaction) method, wherein upstream and downstream sequences of qPCR detecting primer pairs of 16 genes are shown as SEQ ID No: 1 to 32; thirdly, analyzing qPCR results to obtain expression results of the 16 genes treated by the abamectin with different concentrations at different time periods.

The invention relates to a kit for detecting effectiveness of oxaliplatin to colorectal cancer. The kit comprises a pair of amplification primer sequences for detecting MAP4K1 gene copy number, a pair of amplification primer sequences for detecting PIK3CA gene copy number and a pair of primer sequences for detecting reference gene GAPDH gene copy number. Through the kit designed in the invention, effectiveness of oxaliplatin to colorectal cancer can be detected rapidly and effectively, and whether a patient to be detected is suitable to use oxaliplatin as a first-line chemotherapy drug is evaluated. The kit provided by the invention provides effective reference for clinical medication.

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) promoter DNA which induces cell death, and a screening method for an apoptosis-suppressing agent using it. According to this invention, an apoptosis-suppressing agent can be screened by measuring the promoter activity of the GAPDH gene which discovers specifically the GAPDH protein which guides cell death in cell death process using the host cell in which the transformation was carried out by the vector incorporating the GAPDH promoter DNA.

The invention discloses a kit for rapidly detecting Ebola virus and aims to provide the kit which is simple in application method and can be used for rapidly detecting the Ebola virus. Technical points are that the kit is prepared by the following steps sequentially: (1) designing a reverse transcription primer, an amplification primer and a probe in a conserved region of a Nucleoprotein gene; (2) designing the reverse transcription primer and the probe which are used simultaneously in the conserved region of a GAPDH gene; (3) preparing an RT-TaqMan kit. The kit belongs to the technical field of gene detection.

The invention discloses a two-color fluorescent quantitative co-amplification polymerase chain reaction kit. Firstly, the specific primers and two-color Taqman probes are respectively synthesized according to the specific DSCR segment sequence of No. 21 chromosome and the GAPDH gene sequence on No. 16 chromosome, and then In the same amplification buffer reaction system, after 45 PCR cycles, the two pairs of primers are simultaneously guided to synthesize new strands, and the co-amplification of the two pairs of primers is realized. During the PCR amplification, the intensity change of the fluorescent signal obtained due to the degradation of the Taqman probe was detected during the amplification process, and the respective amplification curves were obtained, and then the Ct value of each gene was obtained. Thereby calculating their copy number ratio, and then further judging the ploidy of chromosome 21 according to the ratio of the copy numbers of the two genes.

The invention discloses a special kit for detecting mRNA of a gene Bmi-1 in blood plasma of patients with cervical cancer, which has the advantages of stable result, reliable analysis and the like. The special kit comprises primers of the mRNAs of the gene Bmi-1, a gene GAPDH, a gene EEF1A1, which are shown by SEQUENCE LISTING. The invention also discloses a method for detecting the expression ofthe mRNA of the gene Bmi-1 in blood plasma of the patients with cervical cancer, which has the advantages of simple, convenient and easy operation and the like. The method comprises the following step: (1) extracting total RNA of sample blood plasma; (2) performing reverse transcription-polymerase chain reaction (RT-PCR) amplification; (3) drawing a standard curve, calculating slope, and calculating amplification efficiency E according to a formula (E=10(-1 / slope)-1; and (4) calculating, namely selecting sample and calibrating sample detection holes, obtaining a sample threshold and a calculating sample threshold by using random software of real-time fluorescence quantitative PCR apparatus, obtaining a calibration initial copy number Q of the gene according to a formula (Q=(E+1)-deltaCq),and obtaining the relative expression of the target gene Bmi-1 by comparing the geometrical average of the Q values of the sample target gene and the geometrical average of the Q values of an internal reference gene.

The present invention provides sisal ACT and GAPDH gene sequences, which can be used as sisal internal reference genes, and simultaneously provide sisal ACT and GAPDH as specific primers for internal reference genes, and establish a qRT-PCR method based on SYBR Green fluorescent dye technology, for Subsequent use of real-time fluorescent quantitative PCR provides stable and reliable internal reference genes and technical methods for gene expression analysis under stress conditions such as Phytophthora sisalum infection, low temperature stress, salt stress, and wound treatment.

The invention discloses a gene chip for detecting human papillomavirus (HPV), comprising a solid carrier and a human papillomavirus detecting probes fixed on the solid carrier. The detecting probes comprise nucleotide sequences in SEQ ID No.9-37, and the 27 types of HVP subtype specific probes in the invention correspondingly comprise 22 kinds of high-risk and mediate-risk HPV, and 5 kinds of low-risk HPV and two kinds of high-risk HPV integration (HPV16 / HPV18). The invention also discloses a detecting kit for detecting various subtypes of HPV in a sample with high speed and high flux, and the kit comprises the gene chip, a primer sequence for detection, a human GAPDH gene, special HPV lysing solution, PCR reaction liquid and a color reagent, wherein the detecting probe fixed on the gene chip has the nucleotide sequence in the SEQ ID No.9-37; the primer sequence for detection is used for amplifying the DNA sequence of the HPV in a clinical sample, and the human GAPDH gene is used for amplifying the clinical sample. The HPV detection has important significance for the clinical diagnosis of HPV as well as the early prevention, the diagnosis, the treatment and the postoperation tracking of cervical cancer.

The invention discloses a method for preparing GAPDH (reduced glyceradehyde-phosphate dehydrogenase) transgenic cotton. GAPDH proteins are as shown in a formula (1) or (2), wherein the formula (1) represents proteins as shown in SEQ ID No.2; (2) the formula (2) represents proteins which are obtained by substitution and / or deletion and / or addition of one or a few of amino acid residues of an amino acid sequence as shown in the SEQ ID No.2. According to the invention, the GAPDH transgenic cotton is obtained for the first time, a foundation is established for cultivation of saline-alkaline tolerance cotton variety and improvement of saline-alkaline tolerance capacity of the cotton, and economic and social benefits are achieved.