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.
出处
《现代生物医学进展》
CAS
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)
关键词
三叶因子1
基因重组
毕赤酵母
表达载体
质粒构建
Human trefoil factor 1
Genetic recombination
Escherichia coil
Pichia pastoris
Expression vector
Constructionof plasmid
