柞蚕凝集素ApIML基因的克隆与表达  

Cloning and Expression of ApIML Lectin Gene from Antheraea Pernyi

作　　者：赫健 温志新 李亚洁 米锐 陈星合 王林美 HE Jian;WEN Zhixin;LI Yajie;MI Rui;CHEN Xinghe;WANG Linmei(Liaoning Ocean and Fisheries Science Research Institute,Dalian Liaoning 116023,China)

机构地区：[1]辽宁省海洋水产科学研究院,辽宁大连116023

出　　处：《北方蚕业》2022年第4期1-8,共8页North Sericulture

基　　金：广东省农产品加工重点实验室开放课题(201604);国家茧丝绸发展专项资金项目(201402);财政部和农业农村部现代农业产业技术体系项目(CARS-18)。

摘　　要：在昆虫的免疫防御系统中,凝集素介导的免疫反应起着重要作用。本文克隆了柞蚕凝集素蛋白基因ApIML,序列分析表明:ApIML含有927bp的开放阅读框,编码309个氨基酸。柞蚕凝集素属于分泌型蛋白质,N端有21个氨基酸组成的信号肽,含有2个糖识别结构域。系统进化分析显示ApIML与二化螟Cs-1属于同一个分支,序列相似度为70.16%。构建了ET28a-ApIML原核表达载体,经SDS-PAGE检测发现蛋白在BL21(DE3)中成功表达,并且在Ca存在时对大肠杆菌和金黄色葡萄球菌有凝集作用。Immunoreaction mediated by lectins plays an important role in insect immune defense system. In this experiment, the ApIML gene from Antheraea pernyi was cloned, and sequence analysis indicated that ApIML has an open reading frame of 927 bp and codes for 309 amino acids. ApIML is a secreted protein with a signal peptide consisting of 21 amino acids at the N-terminus and contains two carbohydrate recognition domain(CRD) which has strong binding ability to glycoprotein and glycolid on the surface of the pathogen. Phylogenetic analysis showed that Cs-1 belonged to the same clade, and had a sequence identity of 70.16%. The prokaryotic expression vector pET-28 a/ApIML was constructed. SDS-PAGE analysis showed that ApIML was expressed successfully in BL21(DE3), and it can agglutinate E. coli and S. aureus in the presence of Ca.

关 键 词：柞蚕 凝集素基因 克隆 表达 

分 类 号：S885.1[农业科学—特种经济动物饲养]