人亲环素B基因的克隆表达及重组蛋白的生物活性

Cloning,Expression of the Human Cyclophilin B Gene in Escherichia coli and Purification and Characterization of Recombinant Protein

应用ＲＴ－ＰＣＲ方法从人淋巴细胞中扩增出亲环素Ｂ（ｃｙｃｌｏｐｈｉｌｉｎＢ，ＣｙＰＢ）基因，克隆入ｐＥＴ－２８ａ载体中表达．表达产物以包涵体形式存在，占细菌可溶性蛋白的１５％．经Ｎｉ－ＮＴＡ树脂金属螯合亲和层析和ＳｅｐｈａｄｅｘＧ－５０柱层析纯化，ＳＤＳ－ＰＡＧＥ检测呈单一条带，毛细管电泳为单一色谱峰，纯度达９５％．经复性处理。

The cDNA encoding human cyclophilin B(CyPB)from human lymphocytes was amplified by reverse transcription polymerase chain reaction and after sequencing it was cloned into the expression vector pET 28a.The fusion protein was expressed in inclusion body at 15% of soluble bacteria protein.Recombinant protein was purified by Ni NTA metal chelate affinity and Sephadex G 50 chromatography.The purified CyPB presented only one band on the SDS PAGE pattern and a single peak in the capillary electrophoresis with more than 95% purity.After renaturation,it showed peptidyl prolyl cis trans isomerase activity.