奶牛S100A12基因克隆及其在乳腺和生殖系统中的表达

摘要

为了解奶牛S100A 12基因序列的特征及其在乳腺和生殖系统中的表达情况，采用RT-PCR技术从奶牛外周血液中扩增S100A 12基因，重组到pMD19-T Simple载体中，进行了测序和序列分析，并应用RT-PCR技术检测了奶牛乳腺和生殖系统（卵巢、输卵管、子宫颈、子宫体、阴道等组织)中S100A12 mR-NA的表达分布。结果显示，克隆的S100A 12基因包含一个279 bp的完整开放阅读框（ORF)，与GenBank中的奶牛S100A12基因（NM 174651)编码区序列100%相同，该ORF编码的92个氨基酸含有一个EF手型结构（位于49-84氨基酸残基)；另外，还预测到3个蛋白激酶C磷酸化位点，分别位于19-21、44-46和70-73氨基酸残基。S100A 12基因在奶牛乳腺和生殖系统上皮组织中均有表达。推测S100A 12基因在奶牛乳腺和生殖系统中具有一定作用。%To study the characteristics of dairy cow S100A 12 gene and its expression in lacteal gland and genital system, the S100A 12 gene was amplified by RT-PCR from peripheral blood of dairy cow and sequenced and analyzed after recombina-tion in pMD19-T Simple vector. The expression distributions of S100A 12 mRNA in dairy cow’s lacteal gland and genital system were detected by RT-PCR. The results showed that the cloned S100A 12 gene had a 270bp ORF, 100% identical with S100A 12 gene in GenBank. In ORF, there was a EF hand formation consisted of 92 amino acids (49-84 amino acids residue﹚. Three phosphorylation sites of protein kinase C were found in 19-21, 44-46, 70-73 amino acid residues. S100A 12 gene was expressed in all epithelial tissue of dairy cow lacteal gland and genital system. It might play an important role in dairy cow lacteal gland and genital system.