牦牛BMP7基因的克隆及其在发情期卵巢、子宫的表达分析

Cloning of Yak BMP7 Gene and Its Expression in Ovary and Uterus during Estrus

为探究牦牛骨形态发生蛋白(Bone morphogentic protein 7,BMP7)生物学特性和对雌性牦牛生殖生理的调控作用。研究基于牦牛BMP7基因(NW_005392954.1)设计引物,以发情期牦牛的卵巢、子宫的cDNA为模板进行扩增,借助DNAMAN 8.0等软件、NCBI等网站对扩增产物进行生物信息学分析。通过western blot和qRT-PCR对牦牛的卵巢、子宫的BMP7蛋白和mRNA进行定量分析,结果表明:扩增的BMP7基因为428 bp,编码142个氨基酸,其基因序列与水牛、奶牛和山羊等反刍动物的同源性均可达到98.59%,且在0~29位氨基酸存在信号肽,属于分泌型蛋白。western blot和qRT-PCR结果显示发情期卵巢表达量显著高于子宫(P<0.05)。研究结果表明,BMP7可能参与调控牦牛的繁殖性能,为牦牛在高压低氧环境下生殖生理机制方面的研究提供参考。

To investigate the biological characteristics of Bone morphogentic protein 7(BMP7)and its regulation on reproductive physiology of female yaks,primers were designed based on yak BMP7 gene(NW_005392954.1),the cDNA of ovary and uterus of yak in estrus were used as templates for amplification,and the bioinformatics analysis of amplification products was carried out with the help of DNAMAN 8.0 and NCBI websites in this study.Western blot and qRT-PCR were used to quantitatively analyze BMP7 protein and mRNA in ovary and uterus of yaks.The results showed that the amplified BMP7 gene was 428 bp,encoding 142 amino acids.The gene sequence of BMP7 was 98.59%homologous to that of buffalo,cow,goat and other ruminants,and there were signal peptides at positions 0-29 of amino acids,which belonged to secreted protein.The western blot and qRT-PCR results showed that the ovarian expression was significantly higher than that in uterus during estrus(P<0.05).The results of this study suggest that BMP7 may be involved in the regulation of reproductive performance of yaks,and provide reference for the research of reproductive physiological mechanism of yaks under high pressure and low oxygen environment.