Cloning Expression And Polymorphisms Analysis Of BMP4 And BMPR-IB Genes Associated With Reproductive Traits In Qianbei Ma Goat

The Qianbei Ma goat was treated as test subject,BMPR-IB and BMP4 were candidate genes in this study.We tested polymorphismof BMPR-IB and BMP4 genes and analyzed the correlation between their polymorphism and reproductive traits.The tissue expression distribution was also detected in single and more lamb group of Qianbei Ma goat.The CDS region and prokaryotic expression vector of PET32a(+)-BMPR-IB/BMP4 about BMPR-IB and BMP4 genes were cloned and constructed,respectively.The western-blot technique was used to detect the expression of BMP4 recombinant protein.The study is aimed to explore the effect of BMPR-IB and BMP4 gene on litter size and further enrich these genes related to reproductive traits of Qianbei Ma goat.1.The polymorphism detection of BMPR-IB and BMP4 genesThe C40 G mutation was detected in exon 9 of BMPR-IB gene in Qianbei Ma goat and Guizhou Black Goat,none of polymorphism of BMP4 gene was detected in two goat breeds.Futhermore,the result of association analysis between different genotypes at C40 G loci of BMPR-IB gene and reproductive traits showed significant difference was found among individuals of Qianbei Ma goat.No difference was detected among three genotypes of Guizhou Black Goat.2.The mRNA expression of BMPR-IB and BMP4 genes in breeding tissueThe mRNA of BMPR-IB gene was expressed in detected all 5 tissues including pituitary,fallopian tubes,uterus,hypothalamus and ovaries for single lamb group and more lamb group of Qianbei Ma goat,and the expression trend was consistent in the two groups followed by ovaries,pituitary,uterus,hypothalamus,fallopian tubes fromhigh to low.Moreover,the mRNA expression of BMPR-IB gene in ovaries of 2groups achieved extremely significant difference level(P<0.01),and no significant difference were found among other tissues(P>0.05).For BMP4 gene,it was detected in the same 5 breeding tissues of two groups of Qianbei Ma goat,and high expression in the ovaries but low in fallopian tubes.The expression of BMP4 gene reached significant difference level in ovary tissue of two groups and the expression of more lamb group was significantly higher than that of single lamb group(P?0.05)..3.The CDS region clone and prokaryotic expression vector construction of BMPR-IB and BMP4 geneWe cloned CDS region of BMPR-IB and BMP4 genes successfuly and digested the CDS region and PET32a(+)empty vector by EcoR I and Xho I enzymes,constructed prokaryotic expression vector PET32a(+)-BMPR-IB/BMP4.The result of sequencing showed cloned gene was target BMPR-IB and BMP4 of Qianbei Ma goat.Western-blot verificated BMP4 recombinant protein from strain BL21(DE3)was expressed successfully.