人雄激素受体LBD基因融合表达产物的分离制备

利用硫氧还蛋白事例表达系统，将编码人雄激素受体（ｈＡＲ）激素结合区（ＬＢＤ）的基因（编码ｈＡＢ５８４－９１８氨基酸）插入质粒ｐＴｒｘ Ｆｕｓ中ｔｒｘＡ基因的３′端，重组后的质粒ｐＴｒｘＡＲ导入Ｅ．ｃｏｌｉ ＧＩ７２４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Ｓｅｐｈａｄｅｘ Ｇ－２００柱层析都可较好地纯化分离该融合表达产物（达到ＳＯＳ电泳单点纯），

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

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.