人孕酮受体激素结合区基因表达产物的检测与制备

DETECTION AND PREPARATION OF LBD SEGMENTS OF hPR EXPRESSED IN E. COLI

PuPR为含有编码人孕激素受体（hPR）激素结合区（LBD）基因（631-933氨基酸）的pUC19重建质拉，在其宿主Escherichiacoli（BL21）中，经1mmol/L的IPTG诱导，该转化的细菌产生一C端为6×His结尾的蛋白质，分子量45KD。该表达产物可利用Ni-NTAHRP在ng水平用Western印迹检测，并可通过亲和层析的方法，利用His－bindingresin（Ni树脂）从菌体的裂解液中方便地回收到该蛋白质。

A DNA fragment encoding the human PR LBD (amino acids 631 -933) was cfoned into the pUC19 to form a reconstructed plasmid PuPR. In Escherichia coli (BL21), whan induced with 1mmol/L IPTG, the transformed bacteria expressed a 6×His-tagged protein (MW=45KD), which could be detected by Ni-NTA HRP conjugate through Western blot at the ng level.The protein could be easily meovered from the lysate of bacteria through affinity chromatography by means of His-binding resin.