固相磁珠扣除杂交法筛选奶牛Y-X精子差异表达基因  

Differentially expressed genes in X-and Y-sperm from bovine screened by subtractive hybridization

作　　者：马梦婷 王珊珊[1] 陈晓莉[1] 依斯拉穆 高庆华[1] 韩春梅[1] 

机构地区：[1]新疆生产建设兵团塔里木畜牧科技重点实验室,新疆阿拉尔843300 [2]塔里木大学生命科学学院,新疆阿拉尔843300

出　　处：《新疆农业科学》2015年第4期774-778,共5页Xinjiang Agricultural Sciences

基　　金：国家自然科学基金项目(30760170;31260551);塔里木大学校长基金创新群体项目(TDZKCX201201)

摘　　要：【目的】筛选出奶牛Y-X精子差异表达基因。【方法】利用固相磁珠扣除杂交技术分选高纯度奶牛Y精子对X精子的表达基因,获得的差异片段经SMART扩增后构建Y精子对X精子消减杂交c DNA质粒库,克隆测序,在线进行序列同源性分析。【结果】筛选出的4条候选基因均与牛EST数据库中的序列高度同源,其中之一确认为牛Sry基因,其余3条牛未知功能基因。【结论】基于磁珠上的固相扣除杂交技术有利于简单快速的富集差异表达基因,可用于筛选奶牛Y-X精子候选差异表达基因。【Objective】Screening differentially expressed genes in X or Y sperm.【Method】differentially expressed genes in X or Y sperm were screened by subtractive hybridization,get differences fragment,after SMART build a c DNA plasmid library,cloning and sequencing,for sequence homology analysis. 【Result 】There exist 4 bovine ESTs in Y-X subtracted c DNA library. One of them is known in bovine,which is the sequence of sry gene,and three are unknown in bovine.【Conclusion】Our experiments show that the subtractive hybridization technique which based on bead can be used in screening differentially expressed genes in X or Y sperm,and it conducive to quick and easy enrichment of differentially expressed genes.

关 键 词：固相磁珠扣除杂交 奶牛 精子 差异表达基因 

分 类 号：S823[农业科学—畜牧学] S188[农业科学—畜牧兽医]