摘要
BMP-15基因主要在哺乳动物卵巢中表达,对卵泡的发育和分化起重要作用,克隆了牛BMP-15成熟肽编码区的cDNA序列并进行序列分析,旨在为BMP-15在牛繁殖性能方面的进一步研究奠定理论基础。根据其他物种BMP-15基因的保守序列设计特异性引物,扩增牛的cDNA序列。从牛卵巢中提取总RNA,采用RT-PCR技术扩增出牛BMP-15cDNA序列。将此片段克隆到PMD18-T载体中,经菌落PCR鉴定和DNA序列测定分析验证,证实所克隆序列BMP-15为骨形态发生蛋白,符合骨形态发生蛋白基因的特征。序列分析表明,该cDNA包含有1 185 bp组成的开放读码框(ORF),该ORF编码394个氨基酸,与绵羊、猪、人、小鼠等动物氨基酸序列比对发现同源性分别为98.5%,87.6%,74.0%,69.4%。
BMP-15 (Bone Morphogenetic Protein-15)gene has important role in the development and differentiation of follicle and is specialized expressed in mammalian ovaries. In order to lay the theoretical foundations for further study on BMP-15 in bovine reproduction, cDNA sequence of bovine BMP-15 was cloned and analyzed. The specific primers according to conserver sequence of the other animals was designed to amplify bovine BMP-15 cDNA with RT-PCR The purified RT-PCR product was cloned into PMD18-T vector. The results showed that the bovine BMP-15 belonged to the family of bone morphogenetic protein because of an opening reading frame(ORF ). The ORF contained 1 185 base pairs which encoded 394 amino acid pre-pro-peptide and the pre-pro-peptide contained the BMP consensus sequence of six invariantly spaced cysteines residues. Compared with the sequence of sheep, pig, human, mouse and the ORF homology were 98.5 %, 87.6 %, 74.0% ,69.4% respectively.
出处
《华北农学报》
CSCD
北大核心
2007年第4期63-66,共4页
Acta Agriculturae Boreali-Sinica
基金
内蒙古科技攻关项目(20040102)
内蒙古自然科学基金资助项目(200607010410)
关键词
牛
BMP-15
RT-PCR
克隆
序列分析
Bovine
Bone Morphogenetic Protein-15
RT-PCR
Cloning
Sequence analysis