绵羊tnp2基因的克隆及其在体外共培养系统中圆形精子细胞的.PDFVIP

动 物 学 研 究 2009，Jun. 30(3) ：262−266 CN 53-1040/Q ISSN 0254-5853 Zoological Research DOI ：10.3724/SP.J.1141.2009.03262 绵羊tnp2基因的克隆及其在体外共培养系统中 圆形精子细胞的转录分析 白音巴图, 罗奋华, 胡甜园, 侯 越, 吴应积* （内蒙古大学哺乳动物生殖生物学及生物技术教育部重点实验室，内蒙古 呼和浩特 010021 ） 摘要：过渡蛋白2基因（tnp2 ）是圆形精子细胞特异表达的基因。为了开展绵羊圆形精子细胞标记基因的研究， 根据GenBank上已公布的牛的cDNA序列设计引物，采用RT-PCR和分子克隆方法，克隆了蒙古绵羊tnp2基因cDNA 部分编码区序列。DNA 序列测定结果与牛的核苷酸序列比对，同源性为95.3% 。根据绵羊tnp2基因的cDNA序列 设计引物，对共培养的四月龄绵羊睾丸生殖细胞进行RT-PCR鉴定。结果显示体外共培养的绵羊睾丸生殖细胞一 直到第十周后仍有圆形精子细胞产生。绵羊tnp2基因的cDNA克隆和序列测定为进一步研究绵羊精子发生过程奠 定了基础。 关键词：绵羊；tnp2基因；克隆；序列分析；圆形精子细胞 中图分类号：Q593.4 ；Q954.4 ；Q959.842 文献标识码：A 文章编号：0254-5853-(2009)03-0262-05 Cloning of Sheep tnp2 Gene and Transcription Analysis of Round Spermatid Cells in the in vitro Co-culture System * BAI Yin-ba-tu, LUO Fen-hua, HU Tian-yuan, HOU Yue, WU Ying-ji (The Key Laboratory for Mammalian Reproductive Biology and Biotechnology, Ministry of Education, Inner Mongolia University, Inner Mongolia Hohhot 010021, China) Abstract: The transition protein-2 gene (tnp2) have been reported as the round spermatid-specific marker gene. However, the DNA sequence of tnp2 gene in Ovis aries have not been reported. In order to study the sheep round spermatid-specific marker gene, we designed primers according to the reported Bos taurus tnp2 gene cDNA sequences in the GenBank. The partial CDS was amplified by RT-PCR. The PCR fragment was inserted into the T-A cloning vector pMD