摘要
【目的】克隆牦牛(Bos grunniens)精子相关抗原11(Sperm-Associated Antigen 11,SPAG11)基因,并了解其分子结构特征,为进一步研究牦牛SPAG11生物学功能奠定基础。【方法】从牦牛睾丸中提取总RNA,RTPCR扩增并克隆SPAG11C、SPAG11 D和SPAG11E基因,测序后进行生物信息学分析。【结果】克隆出牦牛SPAG11基因3个亚型:SPAG11C、SPAG11 D和SPAG11E,序列大小分别是351,408和261bp,分别编码117,129和80个氨基酸,其中SPAG11 D和SPAG11E序列包含1个完整开放阅读框(ORF),SPAG11C序列包含部分ORF。牦牛SPAG11C、SPAG11 D和SPAG11E核苷酸序列与黄牛(Bos taurus)相应序列相似性最高,而与黄牛β-防御素1相似性较低(<50%)。3个蛋白亚型均具有β-防御素家族基本特性,生物信息学分析显示其均含有磷酸化位点。【结论】成功克隆牦牛SPAG11C、SPAG11 D和SPAG11E基因,明确了其编码蛋白的分子结构特征。
【Objective】This study aimed to clone Sperm-Associated Antigen 11（SAPG11）gene of yak（Bos grunniens）and explore its characteristics of molecular structure for further study on its biological function.【Method】The SPAG11 C,SPAG11Dand SPAG11 Egenes from yak were amplified and tested by RT-PCR technology before being cloned into pMD？19-T Simple Vector for sequence and analyzed by bioinformatics tools.【Result】Three subtypes of the SPAG11 gene of yak were cloned（namely,SPAG11 C,SPAG11Dand SPAG11E）with lengths of 351 bp,408bp and 261 bp and numbers of encoded amino acids of 117,129 and 80,respectively.SPAG11 Dand SPAG11Esequences contained a complete opening reading frame（ORF）,while SPAG11 C sequence contained partial ORF.Phylogenetic analysis showed that SPAG11 C,SPAG11Dand SPAG11 Egenes of yak had the highest similarity with Bos taurus（〉99%）but a lower similarity with the beta-defensin-1gene of Bos taurus（〈50%）.All of them had the basic characteristics of beta-defensin family and contained phosphorylation sites.【Conclusion】SAPG11C,SPAG11 Dand SPAG11E genes of yak were successfully cloned,and their molecular structure characteristics were defined.
作者
徐绮嫔
王寒
李计尚
周金伟
张于
沈留红
余树民
彭广能
曹随忠
XU Qipin WANG Han LI Jishang ZHOU Jinwei ZHANG Yu SHEN Liuhong YU Shumin PENG Guangneng CAO Suizhong(Center of Dairy Cattle Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan 611130, China)
出处
《西北农林科技大学学报（自然科学版）》
CSCD
北大核心
2016年第12期28-34,44,共8页
Journal of Northwest A&F University(Natural Science Edition)
基金
教育部"长江学者和创新团队发展计划"创新团队项目(IRT0848)
国家自然科学基金项目(31172379)
四川农业大学学科建设双支计划项目(3570806)
关键词
牦牛
精子相关抗原11
Β-防御素
生物信息学
yak（Bos grunniens）
Sperm-Associated Antigen 11（SPAG11）
beta-defesin
bioinformatics
