小菜蛾β-1,3-葡聚糖识别蛋白βGRP基因的克隆与表达

发布时间：2018-05-13 06:03

  本文选题：小菜蛾 + β-　； 参考：《福建师范大学学报(自然科学版)》2017年05期

【摘要】：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.
[Abstract]:In order to study the innate immune mechanism of the Plutella xylostella, a beta -1,3- glucan identification protein (beta -1,3-glucan recognition protein, beta GRP) gene was cloned from the larvae of the Plutella xylostella, and the bioinformatics software was used to analyze and predict the physicochemical properties, hydrophobicity and structure of the gene and its encoded protein. At the same time, the system was constructed. In order to study the function of the protein, the gene was expressed in Escherichia coli and the target protein was purified with a nickel column. The experimental results showed that the open reading frame of the sequence was 1287 BP long, 428 amino acids encoded, the theoretical molecular weight of the encoded protein was about 48.13 Ku, the theoretical isoelectric point was 6.18, the hydrophilic protein, and the phylogenetic tree analysis table. The protein is classified as the same big class with silkworm, tussah, vegetable, cotton bollworm, cotton bollworm, worm moth, and Silverworm. The proteins expressed in Escherichia coli are insoluble inclusion bodies. By denaturation of inclusion bodies and purification of nickel columns, a soluble target protein with high concentration is obtained.

【作者单位】： 福建师范大学生命科学学院;
【基金】：福建省教育厅资助项目(JK2012009)
【分类号】：S433.4
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本文编号：1881963