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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.
In order to study the innate immunity mechanism of Plutella xylostella, a β-1,3-glucan recognition protein (βGRP) gene was cloned from Plutella xylostella larvae. Bioinformatics software Gene and its encoded protein physical and chemical properties, hydrophobicity, structure and other aspects of the analysis and prediction, and to build a phylogenetic tree.In order to study the function of the protein, the E. coli expression of the gene, the use of nickel column purification of the target protein.Experiment The results showed that the open reading frame (ORF) of this sequence was 1 287 bp encoding a polypeptide of 428 amino acids. The calculated molecular mass of the predicted protein was 48.13 ku and its theoretical isoelectric point was 6.18. The result of phylogenetic tree analysis showed that the protein With silkworm, tussah, Pieris rapae, bollworm, moth, moth, Fengyingnianchu belong to the same category; E. coli expression of proteins are insoluble inclusion bodies, by inclusion body denaturation and nickel column purification, Get a higher concentration of soluble protein of interest.