小菜蛾β-1,3-葡聚糖识别蛋白βGRP基因的克隆与表达Cloning and Expression of β-1,3-glucan Recognition Protein Gene from Plutella xylostella
陈少威;吴程;蔡斌斌;杨梅;
摘要(Abstract):
为研究小菜蛾的先天免疫机制,从小菜蛾幼虫体内克隆得到一条β-1,3-葡聚糖识别蛋白(β-1,3-glucan recognition protein,βGRP)基因,利用生物信息学软件对该基因及其编码蛋白的理化性质、疏水性、结构等方面进行分析与预测,同时构建系统进化树.为研究该蛋白的功能,以大肠杆菌为宿主表达该基因,利用镍柱纯化目的蛋白.实验结果表明,该序列的开放阅读框长1 287 bp,编码428个氨基酸,编码的蛋白质理论分子质量约48.13 ku,理论的等电点为6.18,为亲水性蛋白;进化树分析表明,该蛋白跟家蚕、柞蚕、菜青虫、棉铃虫、冬尺蠖蛾、粉纹夜蝶归为同一大类;经大肠杆菌表达的蛋白均为不溶性的包涵体,通过对包涵体的变性及镍柱的纯化,得到了浓度较高的可溶性目的蛋白.
关键词(KeyWords): 小菜蛾;β-1,3-葡聚糖识别蛋白;先天免疫
基金项目(Foundation): 福建省教育厅资助项目(JK2012009)
作者(Authors): 陈少威;吴程;蔡斌斌;杨梅;
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