甘蓝夜蛾几丁质酶基因cDNA序列的克隆与序列分析

Molecular cloning and sequence analysis of chitinase cDNA from Mamestra brassicae

贺玉年 王晓云 韩岚岚 樊 东

点击：2015次 下载：0次

作者单位：东北农业大学农学院 哈尔滨 150030

中文关键词：甘蓝夜蛾， 几丁质酶， 分子克隆， 序列分析

英文关键词：Mamestra brassicae， chitinase， molecular cloning， sequence analysis

中文摘要：

 昆虫几丁质酶在害虫生物防治中具有很大的发展潜力。以甘蓝夜蛾Mamestra brassicae

  L.预蛹期幼虫整个虫体为材料提取总RNA，利用RTPCR和cDNA末端快速扩增技术（RACE）

，扩增得到其几丁质酶的cDNA序列。该序列含有2 826个碱基，包括1个1 689个

碱基的开放阅读框，预测编码1个含562个氨基酸的多肽，分子量约为62.6 kDa，等电点为

5.30。推导得到的氨基酸序列含有2个N-位糖基化位点，22个O-位糖基化位点，氨基酸序

列与其他昆虫，尤其是鳞翅目昆虫的几丁质酶高度同源。获得的甘蓝夜蛾几丁质酶基因cDNA

序列已经登录GenBank并获得登录号FJ436415。

 

 

英文摘要：     Insect chitinase has great potential in biological control of insect pests. Total RNA was isolated from the whole body of Mamestra brassicae L. at prepupal stage. The cDNA sequence was cloned by RTPCR and rapid amplification of cDNA ends was conducted. The cDNA of 2 826 base pairs in length contained an open reading frame of 1 689 base pairs coding for a polypeptide of 562 amino acid residues with a predicted molecular weight of 62.6 kDa and pI 5.30. The deduced amino acid sequence of M. brassicae chitinase was predicted using the software Prosite, which indicated that it has two putative N-glycosylation sites and twenty two putative O-glycosylation sites. The predicted protein shared extensive similarites with those from other insects, expecially the lepidopteran insects. The cDNA sequence has been deposited in GenBank with accession No. FJ436415. 

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