Study On Sex-identification Of Ruminant Using SRY And Microsatellite

This experiment took the sex-identification as the research content, it was for the purpose of establishing the sex-identification procedure and enhanced the accurate rate of sex-identification, enabled the technology of sex-identification to achieve application stage, realized sex control. The experiment took the ruminant including Liangshan Semi-fine -Wool Sheep (20 individuals) and Xinjiang fine -Wool Sheep (4 individuals) and goat (5 individuals) and cattle (6 individuals) as the research object. The technique of multiple PCR was used to amplify the microsatellite markers MILVET09 and AE25 on X chromosome and MCM158 and MAF45 on Y chromosome and SRY gene to identify sex by genotyping, attempted to provide simultaneously the sex-identification and genotyping information in one DNA amplification.The experimental result indicated that the designed primer of SRY gene with highly specificity was the main basis of the sex-identification, but the specificity of the primers of MCM158 and MAF45 was not good, therefore the two markers were not applied to identify sex. The primers of two markers MILVET09 and AE25 on X chromosome only were able to further confirm the examined male individuals. Conclusion: In all examined individuals which could simultaneously amplify SRY, MCM158. MAF45 on Y chromosome and MILVET09, AE25 on X chromosome , also the genotype of MILVET09 and AE25 is homozygous, are normal male, if some examined individuals with heterozygous genotypes for MILVET09 and AE25 were found, it showed that the sex chromosome of the examined individuals were abnormal, could not be used as breeders, in this experiment the male experimental animal did not exist this abnormal situation; the examined individuals only had the amplified production of MCM158, MAF45 on Y chromosome and MILVET09, AE25 onX chromosome, the amplified production of SRY did not occur, then the examined individual was female, furthermore MILVET09 and AE25 did not influence the sex determination of the female. The genotype of MCM158 and MAF45 did not affect the result of the sex-identification of each individual.In this research, although the selected polymorphism microsatellite markers cannot alone be used to identify sex, the X-Y specificity fragments amplified in the past lacked polymorphism, could not provide the information of individual differences. In this experiment, we used the multiple PCR to amplify the selected SRY and 4 polymorphic loci on X and Y chromosome , we can say that it was a significant attempt. Because this established method in the research not only can identify sex but also be used in identification, it had essential difference with the traditional sex inspection, this will have broad application prospect in sports competition, financial insurance fields and so on in future. Furthermore, the method also can be used to monitor the animal meat product, sex-differentiation and sex hypoplasia, desease sieving, identification and paternity test of the forensic medicine, archaeology as well as detection and solve all kinds of serial and concurrent criminal case and so on the fileds.