高原牦牛SRY基因的克隆与原核表达  

Cloning and Prokaryotic Expression of SRY Gene in Plateau Yak

作　　者：裴杰[1] 阎萍[1] 程胜利[1] 褚敏[1] 梁春年[1] 郭宪[1] 曾玉峰[1] 包鹏甲[1] 冯瑞林[1] 朱新书[1] 

机构地区：[1]中国农业科学院兰州畜牧与兽药研究所,甘肃兰州730050

出　　处：《江苏农业学报》2010年第1期107-112,共6页Jiangsu Journal of Agricultural Sciences

基　　金：中央级公益性科研院所基本科研业务费专项资金项目(BRF070105)

摘　　要：为从分子水平了解牦牛的性别形成机制,对高原牦牛SRY(Sex-determining region on the Y chromosome,SRY)基因的编码区进行了克隆表达。以雄性高原牦牛血液为材料,从基因组DNA中扩增SRY基因编码区(单外显子)序列,将其克隆至pGEM-Teasy载体并测序;将牦牛SRY基因编码区连接至pET-28a(+)载体,构建表达载体pET-28a/SRY;把表达载体pET-28a/SRY转入大肠杆菌E.coliBL21(DE3)中,在合适的条件下诱导表达;对表达产物进行Western-blot检测。结果表明:牦牛SRY基因(GenBank:EU547257)编码区长687 bp,编码229个氨基酸;成功构建了表达载体pET-28a/SRY,并且SRY蛋白得到了大量表达;Western-blot进一步验证了其表达成功。Aiming at d!scovering the mechanism of sex determination in yak, the SRY（ Sex-determining region on the Y chromosome,SRY） gene was cloned and expressed. Taking the blood of plateau yak as material, the SRY gene was amplified from yak＇s genome. The product was cloned to pGEM-T easy vector and was sequenced. The coding region of yak＇s SRY gene（ only one exon） was cloned to EcoR I and Sal I sites of pET-28a（＋） vector to construct an expression plasmid pET- 28a/SRY. The expression plasmid was transformed to E. coli BL21 （ DE3 ） and induced under proper conditions. To be sure that the SRY protein of yak was expressed, the product of expression was identified by Westeru-blot. The total coding region of yak＇s SRY gene （GenBank ： EU547257 ） is 687 bp, encoding a peptide of 229 amino acid residues.

关 键 词：牦牛 SRY基因 克隆 表达 

分 类 号：S823.85[农业科学—畜牧学]