Renalase基因的克隆及表达

Cloning of Human Renalase Gene and its Expression in Escherichia Coli

目的:克隆与表达人Renalase基因。方法:提取总RNA,RT-PCR获得Renalase cDNA片段,克隆到表达载体pET42b(+)中,转染大肠杆菌BL21,采用异丙基硫代半乳糖苷诱导,获取融合蛋白,亲和层析纯化后,将产物进行聚丙烯酰胺凝胶电泳与蛋白质印迹分析。结果:克隆到大小为1 029 bp的Renalase cDNA片段,表达载体pET42b(+)-Renalase经测序证实为所需质粒。获得的融合蛋白经鉴定证实为重组Renalase蛋白。结论:成功对Renalase基因进行克隆与表达,为进一步研究奠定了基础。

Objective：To clone and express human renalase gene. Methods：Total RNA was extracted in human kidney and the cDNA of renalase was obtained by rcverse- transcription PCR. The renalase cDNA was inserted into the expression vector pET42b（ ＋ ）and was transformed into E coil BL21. The fusion protein was induced by IPTG and purified by affinity chromatography. The products were identified by SDS- PAGE and Western blot analysis. Results：The renalase cDNA was obtained and was 1 029 bp. The renalase fusion protein was obtained, whose molecular weight was 38 kD. Conclusion： The human renalase gene was cloned and expressed in E coli successfully, which paved the way for further research of the renalase function.