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

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.