GST-hDaxx蛋白的构建及其原核表达产物的鉴定
Identification of GST-hDaxx Protein Construction and Its Prokaryotic Expression Products
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摘要: 目的 构建GST hDaxx蛋白并在原核细胞中诱导表达GST hDaxx融合蛋白, 并研究其与 p5 3的结合能力。方法 构建GST hDaxx融合蛋白原核细胞表达载体pGEX 4T/hDaxx, 在大肠杆菌 (E .col i)中用异丙基 β D 硫代半乳糖苷 (IPTG)诱导表达。表达产物用亲和层析柱加以纯化后, Westernblot鉴定表达产物与纯化物。通过共免疫沉淀反应与Westernblot观察hDaxx与p5 3的结合反应。结果 在原核细胞中成功表达了GST hDaxx融合蛋白, hDaxx与 p5 3可发生共免疫沉淀反应。 结论 1.GST hDaxx融合蛋白成功表达 ;2 .hDaxx和 p5 3的相互结合提示它们可能与肿瘤的发生有关。Abstract: Objective To construct a recombinant protein of GST-hDaxx and to induce the expression of its fusion protein. Methods By constructing pGEX-4T/hDaxx recombinant, GST-hDaxx fusion protein was induced to express by IPTG in E.coli BL21, and purified by glutathione resin. hDaxx fusion protein was analyzed by Western blot. The direct binding of hDaxx and p53 was studied by coimmunoprecipitation reaction. Results hDaxx fusion protein was induced to express by IPTG in E.coli successfully. Soluble GST-hDaxx fusion p...
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