Snps Detection And Different Tissue MRNA Expression Of Three Sheep Breeds BMP-15 Gene And Correlation Analysis With The Lambing Trait

This study adopted PCR-SSCP technology and sequencing technology to carry out SNPs detection and correlation analysis between two exons of BMP-15 gene of different lambing performance sheep,then adopted the real-time fluorescent quantitative PCR technology to BMP-15 gene in sheep m RNA expression in different tissue volume lambing trait and correlation analysis.To further study the effects of BMP-15 gene on t on fecundity of sheep.The main results are as follows:1.The correlation analysis of the first exon polymorphism and lambing traits of sheep BMP-15 gene.Produced single lamb Mongolia sheep,double lamb Mongolia sheep,single lamb gansu alpine fine wool sheep,double lamb gansu alpine fine wool sheep and small tail Han sheep three different varieties of reproductive performance of five groups as the research object,adopted the PCR-SSCP technology combined with sequencing technology,the method of detection of BMP-15 coding regions 67 and 92 in five sheep populations in presence of Fec XH mutations and Fec XI mutations,then according to the test results with lambing trait for correlation analysis.The results show that neither of the three varieties detected the same Fec XI mutation as the Inverdale sheep,nor did they detect the same Fec XH mutation as that of the Romney sheep.It was speculated that the SNP loci of the BMP-15 gene affecting the lambing traits of the Romney and Inverdale sheep did not affect the lambing traits of the above-mentioned sheep varieties.2.The correlation analysis of the second exon polymorphism and lambing traits of sheep BMP-15 gene.Adopted PCR-SSCP to detect polymorphisms in exon two,locus 747th(T747?C)and 755th(T755?C)in Mongolia sheep,Gansu Alpine Merino sheep and Small Tailed Han sheep of BMP-15,and also studied the effect of two mutations on three kinds of sheep lambing number.The result showed that:(1)There are three types of genotypes including wild homozygous AA,mutant heterozygous AB and AC.AA was the dominant genotype and A was the dominant allele;(2)These three genotypes were detected in the Gansu Alpine Merino sheep,Merino sheep,but the AB genotype was not detected in Mongolia sheep and Small Tail Han sheep;(3)The ewes with heterozygous mutant AB had 0.27(P<0.05)lambs more than those with wild homozygous type AA in Mongolian sheep;(4)The genotype frequency of AC in double lambing ewes and multiparous ewe were higher than that of single ewe lambing ewes.According to the above experimental results,we suggest that the mutation at BMP-15 755 th site(AC)has a significant effect on Mongolia sheep twin lambs,we also found a tendency that the litter size of sheep which have AC genotype was more than that have AA and AB genotypes,and this locus may be a potential DNA marker for affecting prolificacy in sheep.3.Analysis and differences in the expression of different lambing traits ewes BMP-15 geneIn order to found out the differences in the expression of lambing traits ewes BMP-15 gene(bone morphogenetic protein 15,BMP-15)in different tissues and its analysis with litter size traits correlation,adopted real-time quantitative PCR and reverse transcription PCR to detect the distribution of BMP-15 in five tissues of hypothalamus,pituitary,ovary,kidney and heart among Mongolian sheep,Poll Dorset and Small Tail Han Sheep.The results showed that:(1)The BMP-15 gene was expressed in the kidneys and ovarian tissues of the Mongolia sheep double lamb and the Poll Dorset sheep,and the expression in the renal tissue was significantly different from that of the ovarian tissue(P<0.05),but the relative expression of the gene was very low in the other organs;(2)In the Poll Dorset sheep and Mongolia sheep,the expression of BMP-15 m RNA in the ovary was significantly higher than that in the single lamb ewe(P<0.05),and the relative expression level was significantly higher in the ovary tissues of the Small Tail Han Sheep than the Mongolia lamb single and double lamb ewe(P<0.05).It is indicates that m RNA of the transcription of BMP-15 gene varies greatly in ovary tissues of sheep with different lambing traits.It is speculated that:(1)The transcription level of BMP-15 gene has a significant effect on the lambing behavior of sheep;(2)The BMP-15 gene may be one of the candidates for the high productivity of the three varieties of sheep.