红笛鲷NCCRP-1基因原核表达条件的优化及纯化Purification and Optimization of Prokaryotic Expression of Crimson Snapper (Lutjanus sanguineus) NCCRP-1 Gene
魏世娜,王蓓,鲁义善,吴灶和,简纪常
摘要(Abstract)：
通过克隆编码红笛鲷(Lutjanus sanguineus)非特异性毒性细胞受体蛋白-1(nonspecific cytotoxic cell receptor protein-1,NCCRP-1)成熟肽基因序列,然后与载体连接构建pET21a-NCCRP融合蛋白表达载体,将其转入大肠杆菌BL21进行诱导表达并优化条件。SDS-PAGE分析表明,在异丙基-β-D-硫代半乳糖苷(IPTG)浓度0.8 mmol/L、37℃条件下培养3 h后表达量最大,分子大小与预期值相符,融合蛋白主要以包涵体形式高效表达,通过HisTrap HP柱子使其得到进一步纯化;Western blot分析表明,该融合蛋白可与鼠抗His-tag单克隆抗体发生特异性结合,说明获得该表达产物。
关键词(KeyWords)： 红笛鲷;NCCRP-1基因;原核表达;条件优化;纯化
基金项目(Foundation): 国家自然科学基金“红笛鲷仔鱼抗菌免疫基因的克隆及表达时序研究”(40906073)
作者(Author): 魏世娜,王蓓,鲁义善,吴灶和,简纪常
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