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1185 bp组成的开放读码框(ORF)，该ORF编码39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%，7.0%，69.%。

关键词: 牛, BMP-15, RT-PCR, 克隆, 序列分析

Abstract: 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 1185 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.

Key words: Bovine, Bone Morphogenetic Protein-15, RT-PCR, Cloning, Sequence analysis

中图分类号: 

• S823.03