PGR Mutations Detection By DHPLC And PCR-SSCP In Tibetan Sheep

Fertility is decided to produce one of the main economic characters of sheepindustry.Low heritability, the sheep are lambing number selection in conventional breedingmethods have little effect. So to find its main effect genes and genetic markers is of greatsignificance for improving the sheep are lambing number. The whole animal breeding processincluding reproductive cells and mature, ejaculation, ovulation, pregnancy and lactation andso on each link, each part is controlled by the reproductive hormones. Therefore, coding thesereproductive hormones or its receptor gene is the most ideal candidate genes in reproductiveperformance genetic markers.Progesterone, represented an important steroid hormones play akey role in the maintenance of pregnancy, has been effectively used to prevent recurrentpreterm birth. Progesterone is mediated by the progesterone receptor, PGR as ligand-activatedtranscription factor with DNA sequence-specific binding, enhancing or inhibiting geneexpression, thereby regulating reproductive and physiological processes such asmorphogenesis.This study was to compare DHPLC method and PCR-SSCP method testingthe effect of the same gene mutation and PGR gene polymorphisms of exon4, to exploresuitable for screening gene mutation method in the field of animal breeding and PGR’sphysiological function and its influence on Tibetan sheep fertility provides the basis.With the view of studying polymorphism of PGR exon4in Tibetan sheep was analyzed,determining the number of alleles, nucleotide polymorphism sites, amino acid polymorphismsites, genetic relationship among every allele and evolutionary significance, as well as thepossibility of PGR exon4as the molecular genetic markers. In this study, two methods ofDHPLC and PCR-SSCP were compared to detected PGR exon4mutations in391TibetanSheep. This study aimed to bioinformatics of PGR in ovine. The physicochemicalcharacteristics, structures and functions of ovine PGR were predicted and analyzed withsoftware tools and database. The results showed thatthe polymorphism of exon4of PGR genein Tibetan Sheep wasdetected in this experiment.Therewere three genotypes named AA、ABand BB in Tibetan Sheep,AA was preponderant genotype,allele A was preponderant allele.Theresult of χ2independencetest indicated that the distribution and constitution of genotypes atthe exon4of PGR gene showed significant differences in Tibetan Sheep (P<0.01).Conclusion,SSCP method of Tibetan sheep populations PGR gene mutation detection rate higher than that of DHPLC method, and the detection of DHPLC method higher accuracy, and from the testmethod of cost and convenience degree of difficulty, degree of automation and operation, etc.,comprehensive evaluation, the PCR-SSCP method more suitable for animal SNPs detection.In addition, Tibetan sheep PGR gene exon exist two SNPs,4belong to low polymorphism.