禾谷孢囊线虫(Heterodera avenae)纤维素结合蛋白基因(Ha-cbp-1)的克隆和序列分析

Molecular cloning and sequencing of cellulose binding protein gene (Ha-cbp-1) from the cereal cyst nematode(Heterodera avenae)

禾谷孢囊线虫(Heterodera avenae)是中国小麦上的重要病原线虫。纤维素结合蛋白基因是一种重要的植物线虫寄生和致病相关基因。用同源克隆的方法从禾谷孢囊线虫寄生前二龄幼虫中克隆出一种纤维素结合蛋白新基因Ha-cbp-1(GenBank注册号GQ178086)cDNA序列。Ha-cbp-1基因cDNA包含1个开放阅读框,编码131个氨基酸残基的蛋白质,预测蛋白由1个长度为18氨基酸残基的信号肽和1个纤维素结合区域(CBD)组成。Ha-cbp-1的基因组DNA序列含有2个内含子。预测的禾谷孢囊线虫纤维素结合蛋白(HA-CBP-1)序列与大豆孢囊线虫纤维素结合蛋白(HG-CBP-1)序列有60%的同一性和76%的相似性,与甜菜孢囊线虫纤维素结合蛋白(HS-CBP-1)序列有60%的同一性和75%的相似性。本研究首次从禾谷孢囊线虫中成功克隆出CBP蛋白基因。

The cereal cyst nematode,Heterodera avenae,is an important nematode pathogen of wheat in China.The cellulose binding protein genes are the important parasitism genes for invasion of plant parasitic nematodes.The cDNA sequence of Ha-cbp-1（GenBank accession GQ178086） was cloned by RACE kit based on homologous cloning method.The results showed that the cDNA sequence of Ha-cbp-1 contained an open rea-ding frame,which encoding 131 amino acids with a predicted signal peptide sequence for secretion and a cellulose-binding domain.The DNA sequence of Ha-cbp-1 contained two introns with the length of 932 bp.The predicted HA-CBP-1 amino acid sequence had 60% identity and 75%-76% similarity with HS-CBP-1 and HG-CBP-1.