张雪利,鲁义善,谢吉国,简纪常,吴灶和

• 1:广东海洋大学水产学院广东省水产经济动物病原生物学及流行病学重点实验室暨广东省高等学校水产经济动物病害控制重点实验室

摘要(Abstract)：

克隆编码红笛鲷(Lutjanus sanguineus)RAG1蛋白(recombination activating protein1)活性核心区的基因序列,并与pET-28a(+)载体连接,构建原核表达载体pET-28a-RAG1,将其转入大肠杆菌BL21(DE3)菌株,利用IPTG进行诱导表达。为提高融合蛋白的表达效率,运用传统的实验方法对诱导条件进行优化。SDS-PAGE分析表明,在37℃条件下,利用0.1 mmol/L IPTG诱导8 h后,RAG1重组融合蛋白的表达量最大,相对分子质量与预测值相符,该蛋白主要以包涵体形式高效表达,利用His Trap HP亲和柱使其得到进一步纯化;Western blot分析显示,该融合蛋白可与鼠抗His-tag单克隆抗体发生特异性结合,说明表达蛋白为目的蛋白。

关键词(KeyWords)： 红笛鲷;rag1基因;原核表达;表达条件;优化;纯化;Western blot分析

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(40906073);; 广东省基金项目(10152408801000006)

作者(Author): 张雪利,鲁义善,谢吉国,简纪常,吴灶和

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