人TRAIL基因的克隆及其在大肠杆菌中的表达

Clone of Human trail gene and its expression in E.Coli

目的克隆人TRAIL基因,构建其原核表达载体。方法采用RT-PCR方法从人宫颈癌细胞系HeLa的总RNA中扩增TRAIL基因114～281片段的cDNA,并将目的片段克隆至原核表达载体pET32a(+)。结果克隆到人TRAIL基因114～281片段的cDNA,DNA测序结果与GenBank基因库报导的完全一致,经SDS-PAGE电泳,可见29kD大小的融合蛋白质表达。结论人TRAIL基因的克隆及其在大肠杆菌中的表达,为进行研究sTRAIL的作用机制提供了实验依据。

[Objective] To clone gene of the TRAIL and construct its prokaryotic expression vector. [Methods] The cDNA from114-281 fragment of human TRAIL gene was amplified from HeLa by RT-PCR and then cloned into the expression vector pET32a（＋）.The TRAIL-GST was expressed in E.Coli. [Results] Sequence indicated that the sequence of TRAIL gene was identical with that in the GenBank. The protein expressed in E.Coh was 29 kD analyzed by SDS-PAGE. [Conclusion] Clone of human trail gene and its expression in E.coli can offer lab data for further researching its mechanism.