奶牛SRY基因核心保守区真核表达载体构建及序列分析  

The Construction of Dairy Cattle SRY Gene Core Conservative Region Prokaryotic Expression Vector and Sequence Analysis

作　　者：岳明星[1] 袁天翔[1] 孔振兴[1] 吕文发[1] 

机构地区：[1]吉林农业大学动物科技学院,吉林长春130118

出　　处：《家畜生态学报》2009年第3期14-16,31,共4页Journal of Domestic Animal Ecology

基　　金：吉林省杰出青年基金(编号:20070133)项目资助

摘　　要：SRY基因是哺乳动物性别分化与控制的主导基因,参与有序的、多层次调控性别分化和控制的过程。本研究从牛血中提取总RNA,设计特异性引物,利用RT-PCR技术扩增出SRY基因HMG box基因片段,将该片段与pMD18-T载体连接并转化到大肠杆菌(E.coliTop)10中,提取质粒并用EcoRⅠ、AvrⅡ酶消化,将消化后的目的基因纯化并回收,与经同样的酶消化后并纯化回收的载体pPIC9K连接并转化到E.coliTop10,获得重组表达质粒pPIC9K-SRYHMG box,经PCR、酶切、测序验证了此载体构建成功,为下一步使用毕赤酵母表达SRY蛋白打下基础。SRY （Sex determine region of the Y chromosome） gene is a key gene which take important part in mammalian sex differentiation and domination. Total RNA was isolated from cattle blood. The tar- get fragment SRY gene HMG box was amplified by RT-PCR and cloned into pMD18-T vector between two restriction enzyme sites （EcoR I and Avr Ⅱ） ,then transferred into E. coli Top10. Further pMD18-T-SRY- HMGbox recombinant plasmid was digested by EcoR I and Avr II and inserted into pPIC9K expression vec- tor. After screening by PCR and restriction analysis,we concluded that the pPIC9K-SRY-HMGbox recom- binant expression plasmid was constructed,this work can be a base of further research.

关 键 词：奶牛 SRY基因 PPIC9K 

分 类 号：S811.5[农业科学—畜牧学]