人雄激素受体LBD基因融合表达产物的分离制备

THE PREPARATION OF hAR LBD SEGMENTS EXPRESSED AS FUSION PROTEIN IN E. COLI

利用硫氧还蛋白融合表达系统，将编码人雄激素受体（hAR）激素结合区（LBd）的基因（编码hAR584－918氨基酸）插入质粒pTrxFus中trxA基因的3＇端重组后的质粒pTrxAR导入E．coliGI724中，经Trp诱导，该转化的菌株表达一融合蛋白。通过制备包含体，再利用制备型SDS-PAGE或SephadexG-200柱层析都可较好地纯化分离该融合表达产物（达到SDS电泳单点纯），为该融合蛋白的特异性酶切裂解和性质分析提供材料。

Whithin the Thioredoxin Fusion Expression system, the E. coli GI724,transformed by pTrxAR which was constructed by connecting a DNA fragment enceding the hAR LBD (amino acids 584~918) to the 3'-terminal of Thioredoxin gene of pTrxFus, expressed a fusion protein when induced with Trp.Through prepndion of an inclusion body and exploitation of SDS-PAGE or Sephadex G200 chromatography,the expressed product could be satisfactorily purified (showing a single line in gel when checked with SDS-PAGE).The purified fusion protein can be used for specific cleavage of enterokinase and its property analysis.