hDaxx与12型腺病毒E1B 55KD癌蛋白在体内外的结合反应  

Binding Reaction of hDaxx with Adenovirus 12 E1B 55kDa Oncproteins in Vivo or in Vitro

作　　者：万艳平[1] 吴移谋[1] 粟盛梅[1] 余敏君[1] 尹卫国[1] 杨长顺[1] 

机构地区：[1]南华大学病原生物学研究所,中国湖南衡阳421001

出　　处：《实用预防医学》2002年第4期293-295,共3页Practical Preventive Medicine

基　　金：湖南省教委重点课题资助 (0 0 A0 0 9)

摘　　要：目的　研究 12型腺病毒 (Adenovirus12 ,Ad12 ) E1B5 5 KD癌蛋白 (Ad12 E1B5 5 KD)打破 h Daxx与急性早幼粒细胞性白血病蛋白 (promyelocytic leukemia protein,PML)在细胞核共定位的机制。　方法　构建 h Daxx原核细胞表达质粒 p QE30 / h Daxx,并在大肠杆菌 (E.coli) BL2 1中诱导表达 ,用镍—氮基三乙酸 (Ni- NTA)琼脂糖加以纯化。通过体内外共免疫沉淀反应 ,Western blot方法研究 h Daxx与 Ad12 E1B5 5 KD直接结合反应。　结果　质粒 p QE30 / h Daxx在E.coli BL2 1中成功诱导表达了 h Daxx,其 N端有 6个组氨酸分子附着 (6 His- h Daxx)。Western blot结果显示 h Daxx在体内外与 Ad12 E1B5 5 KD发生直接结合反应。　结论　表达并获得纯化的 6 His- h Daxx,h Daxx能在体内外与 Ad12 E1B5 5 KD直接结合。Objective To study the direct reason of Adenovirus 12(Ad12) E1B 55KDa oncoproteni (Ad12 E1B 55KDa)disrupting the co-localization with human Daxx(Daxx)in promyelocytic leukemia protein(PML) oncogenic domains(PODs). Methods By constructing pQE30/hDaxx tagged with 6 Histidines, hDaxx was induced with lsopropy1-β-D-thiogalactoside(IPTG) to express in E.coli BL21 and purified by nickei-nitrilo-triacetic acid(Ni-NTA)resin. The binding reaction of hDaxx with Ad12 E1B 55KDa was determinated by coimmunoprecipitation(COIP) and Western blot in vivo or in vitro. Result hDaxx was purified and identified by Western blot. The binding reaction of hDaxx with Ad12 E1B 55KDa in vivo or in vitro was proved by COIP and Western blot. Conclusion hDaxx was induced expression and purified. hDaxx binds to Ad12 E1B 55KDa in vivo or vitro.

关 键 词：HDAXX 12型腺病毒E1B 55KD癌蛋白 共免疫沉淀反应 急性早幼粒细胞性白血病 

分 类 号：R733.71[医药卫生—肿瘤]