hTFF_2毕赤酵母表达载体的构建与鉴定  

Construction and characterization of hTFF_2 expression vector in Pichia pastoris

作　　者：吴炜[1] 孙勇[1] 张勇[1] 吕尚军[1] 彭曦[1] 

机构地区：[1]第三军医大学西南医院全军烧伤研究所,重庆400038

出　　处：《重庆医学》2007年第19期1989-1990,F0002,共3页Chongqing medicine

基　　金：国家自然科学基金(30200294);国家重点基础研究发展计划项目(2005CB522601)。

摘　　要：目的构建三叶因子2(hTFF_2)的毕赤酵母表达载体。方法从人胃窦部提取总RNA,经RT-PCR得到hTFF_2 cD- NA,用PCR方法扩增hTFF_2基因,并将其克隆到毕赤酵母的表达载体pGAPZαA中,构建真核重组表达质粒pGAPZαA-hTFF_2。结果通过双酶切和基因序列分析确定插入pGAPZαA中的片段为hTFF_2基因片段。结论重组质粒pGAPZαA-hTFF_2成功构建,为在真核细胞中高效表达hTFF_2及下一步的功能研究奠定了基础。Objective To construct Pichia pastoris （P. pastoris） expression vector of hTFF2. Methods Total RNA was extracted from human sinus ventriculi, hTFF2 cDNA was isolated by RT-PCR, hTFF2 gene was amplified with PCR and cloned into Pichia pastoris vector pGAPZαA. The expression plasmid pGAPZαA-hTFF2 was constructed. Results The nucleotide sequence of hTFF2 was the same as expected. Conclusion The recombinant plasmid is constructed successful and underlie the base of large scale production and functional analysis.

关 键 词：三叶因子2 基因重组 毕赤酵母 表达载体 质粒构建 

分 类 号：Q78[生物学—分子生物学]