| 高原牦牛SRY基因的克隆与原核表达 |
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| 引用本文： | 裴杰,阎萍,程胜利,褚敏,梁春年,郭宪,曾玉峰,包鹏甲,冯瑞林,朱新书.高原牦牛SRY基因的克隆与原核表达[J].江苏农业学报,2010,26(1). |
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| 作者姓名： | 裴杰 阎萍 程胜利 褚敏 梁春年 郭宪 曾玉峰 包鹏甲 冯瑞林 朱新书 |
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| 作者单位： | 中国农业科学院兰州畜牧与兽药研究所,甘肃,兰州,730050 |
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| 基金项目： | 中央级公益性科研院所基本科研业务费专项资金项目(BRF070105) |
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| 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。
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| 关 键 词： | 牦牛 SRY基因 克隆 表达 |
Cloning and Prokaryotic Expression of SRY Gene in Plateau Yak |
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| PEI Jie,YAN Ping,CHENG Sheng-li,CHU Min,LIANG Chun-nian,GUO Xian,ZENG Yu-feng,BAO Peng-jia,FENG Rui-lin,ZHU Xin-shu.Cloning and Prokaryotic Expression of SRY Gene in Plateau Yak[J].Jiangsu Journal of Agricultural Sciences,2010,26(1). |
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| Authors: | PEI Jie YAN Ping CHENG Sheng-li CHU Min LIANG Chun-nian GUO Xian ZENG Yu-feng BAO Peng-jia FENG Rui-lin ZHU Xin-shu |
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| Institution: | Lanzhou Institute of Animal Science and Veterinary Parmaceutics/a>;China Academy of Agricultural Sciences/a>;Lanzhou 730050/a>;China |
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| Abstract: | Aiming at discovering 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(... |
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| Keywords: | yak SRY gene clone expression |
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