人三叶因子1大肠杆菌及毕赤酵母表达载体的构建与鉴定  

Construction and Characterization of hTFF1 Expression Vector in Escherichia Coli and Pichia Pastoris

作　　者：吴炜[1] 孙勇[1] 吕尚军[1] 张勇[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创伤,烧伤与复合伤国家重点实验室,重庆400038

出　　处：《现代生物医学进展》2007年第8期1138-1141,共4页Progress in Modern Biomedicine

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:构建人三叶因子1(hTFF1)的大肠杆菌及毕赤酵母表达载体。方法:从人胃窦部提取总RNA,经RT-PCR得到hTFF1 cDNA,用PCR方法扩增hTFF1基因,并将其分别克隆到大肠杆菌的表达载体pET32α及毕赤酵母的表达载体pCAPZαA中,构建原核及真核重组表达质粒pET32α-hTFF1和pGAPZαA-hTFF1。结果:通过双酶切和基因序列分析确定插入pET32α和pGAPZαA中的片段为hTFF1基因片段。结论:重组质粒pET32α-hTFF1和pGAPZαA-hTFF1成功构建,为在比较原核和真核细胞中hTFF1高效表达的情况及下一步的功能研究奠定了基础。Objective： To construct Escherichia cofi （E. coli） and Pichia pastoris （P. pastoris） expression vector of hTFF1, and underlie the base of large-scale production and function analyses. Methods： Total RNA was extracted from human sinus ventriculi, hTFF1 eDNA was synthesized by RT-PCR. hTFF1 gene was amplified by PCR and cloned into Escherichia coli and Pichia pastoris vector pET32α and pGAPZαA. The expression plasmid pET32α-hTFF1 and pGAPZαA-hTFF1 were constructed. Results： The nucleotide sequence of hTFF1 was the same as that expected. Conclusion： The successful construction of the recombinant plasmid will be further studied in this field.

关 键 词：三叶因子1 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]