人FKBP12的基因克隆及其表达产物的生物活性

The Gene Cloning and Bioactivity of the Expression Product of the Human FKBP12

为获得具有生物学活性的 h FKBP1 2 ,筛选新型的促神经再生药物 .采用 RT- PCR、计算机辅助 p BV2 2 0中外源基因高效表达的数学模型预测方法及超滤截留纯化方法 ,从人 T淋巴白血病细胞系 Jurkat中成功扩增出 h FKBP1 2基因 .按照外源基因高效表达的数学模型 ,将其进行优化改构后 ,在 p BV2 2 0中实现了高效表达 ,表达量约 2 0 % .重组的包涵体蛋白经复性 ,纯化至电泳纯 ,纯化后的 h FKBP1 2显示出肽基脯氨基顺反异构酶活性 .表明原核表达的 h FKBP1

To obtain bioactive hFKBP12 protein for screening novel neurotrophic drugs,RT PCR,computer aided gene expression discriminating model and M.W.cut off ultrafiltration were used.hFKBP12 gene was cloned successfully and then redesigned optimally according to the high expression model for the foreign gene.The redesigned gene was highly expressed in pBV220 vector as inclusion bodies,which were renatured and purified to only one band on SDS PAGE.The purified hFKBP12 showed peptidyl prolyl cis trans isomerase (PPIase) activity.It indicated that recombinant hFKBP12 was bioactive just as its wild type's.