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2011年5月4日 by admin

stainless steel  Cattle specialized households and cattle farmers are often faced with infertility, only the heat and repeat the cows. Mainly due to the poor quality of bull semen or bull infertility; cow estrus after mating is not timely; cows too fat or too thin, sick and so on. Dealing with causes of infertility cows repeatedly takes appropriate preventive measures. Promotion of artificial insemination could eliminate the bull infertility. At present, China farmers raise cattle, and most of the main natural mating, often due to poor quality of bull semen or bull suffering from infertility that may cause pregnant cows empty. In this study, glutathione S-transferase and cytochrome oxidase for the poor quality of bull semen or bull suffering from infertility candidate gene, SNP screening by both genes and the traits associated with semen quality analysis, mutation Point and method of gene diagnosis of semen quality to the early selection for the breeding bull and semen quality of genetic diagnosis of individuals laid low molecular biology.We choose bull selection of Changchun center 33 kinds of breeding bulls, PCR amplification of glutathione S-transferase gene exons 7 and cytochrome oxidase structural gene with zinc ions in exon 4, PCR-SSCP (using the polymerase chain reaction-single strand conformation polymorphism) detection of the existence of amplified loci in the mutant; and glutathione S-transferase gene and cytochrome oxidase gene structure with zinc ions polymorphic fragments were sequenced to identify mutations in the base type and position of mutation site mutation; combined with determination of semen quality records, Changchun Center of CITIC Group, the bull breeding bull semen quality were analyzed.According to PCR-SSCP and sequencing results of that test: Changchun bull breeding center of CITIC Group, the 33 kinds of bull, in the glutathione S-transferase gene discovered a mutation in the zinc binding of cytochrome oxidase ion structural gene mutations found in three sites.Cloning and sequencing results showed that: in the glutathione S-transferase gene exons, the first exon 5 of the 56bp bit there at the existence of C→T mutation, but because out of the base of thstainless steel e No. 3 in the codon amino acid serine in the mutant is always around. In the cytochrome oxidase gene structure with zinc ions in exon 2, exon 3, were found to have a polymorphism. In the 92bp exon 2 position exists T, C heterozygous genotype; exon 3 107bp position exists T→C mutation, amino acid phenylalanine into a serine.SPSS12.0 software analysised by CITIC Group in Changchun bull breeding center of the 33 kinds of bull glutathione S-transferase gene and cytochrome oxidase structural gene combination of zinc and semen quality of the genetic polymorphism of the correlation between The results show that: For the glutathione S-transferase gene,exon V AA-type activity of fresh semen, semen vitality of live sperm rate of frozen semen to be significantly higher than that of AB and BB-type; the combination of cytochrome oxidase zinc ion structural gene,the second exon on the site and CD-CC,semen between the vitality of live sperm rate of fresh semen, frozen semen sperm live there was a significant difference; the third exon locus EE-type and EF, FF-type activity between the fresh semen, fresh and frozen semen sperm live sperm live sperm rate ratios were significantly different. Therefore, we can glutathione S-transferase gene and cytochrome oxidase structural gene with zinc ion as an important molecular marker for marker-assisted selection bull.PCR-RFLP technique using the genome of a first hypervariable region using two primers specifically amplified PCR products were then digested, and then electrophoresis polymorphism. The results can be seen, the three sites were PCR products were digested with three bands appear. Glutathione S-transferase gene in exon 5 of 57 by the EcoRII restriction enzyme site after the three bands: TT (255bp), CT (255bp, 226bp, 29bp), CC (226bp, 29bp) (which 29bp fragment is small, not shown in the figure); cytochrome oxidase gene binding zinc ion structure of exon 2 polymorphism after digestion by the MaeII three bands: TT (238bp), TC (238bp, 221bp, 17bp) (17bp fragment of which is small, not shown in the figure); cytochrome oxidase gene structure with zinc ions in exon 3 polymorphic site was digested by the SfaNI There are three bands: CC (373bp ), TC (373bp, 286bp, 97bp), TT (286bp, 97bp); consistent with the theoretical expected results.Currently genetic markers on the semen quality to carry out small, so we can glutathione S-transferase gene and cytochrome oxidase structural gene with zincion as an important molecular marker. Marker-assisted selection of these research and development of new semen quality associated with the genetic markers will bull infertility treatment and prevention of laying the basis of molecular biologystainless steel