甜菜夜蛾中肠cDNA表达文库免疫学筛选、克隆及序列分析  

Immune-screening and Sequence Analysis of the Midgut cDNA Expression Library from Spodoptera exigua

作　　者：李雯[1] 郭巍[1,2] 张霞[3] 

机构地区：[1]河北农业大学植物保护学院,河北省农作物病虫害生物防治工程技术中心,河北保定071001 [2]河北农业大学生命科学学院,河北保定071001 [3]河北省科学院生物研究所,河北石家庄050051

出　　处：《华北农学报》2010年第5期33-37,共5页Acta Agriculturae Boreali-Sinica

基　　金：国家自然科学基金项目(30771447);国家重点基础研究发展计划("973"计划)项目(2009CB118902);留学人员科技活动项目

摘　　要：昆虫围食膜是由几丁质和蛋白质组成一个半透膜结构,由中肠分泌,保护细胞免受机械性损伤和细胞微生物感染。以害虫生物防治的新靶标——中肠围食膜作为研究对象,分离和鉴定甜菜夜蛾中肠围食膜蛋白基因。依据免疫学筛选方法,用制备的甜菜夜蛾围食膜蛋白多克隆抗体,对甜菜夜蛾中肠cDNA表达文库进行免疫筛选。获得231个围食膜蛋白阳性克隆,经测序、生物信息学分析可知得到8个几丁质结合蛋白,2个肠粘蛋白;并对结构功能进行分析预测。研究结果为揭示围食膜分子结构,进一步以中肠围食膜为靶标进行生物防治奠定了基础。Peritrophic membrane（PM） is secreted by the lumenal surface of the insect midgut cells and serves to protect cells from mechanical injury or microbial infection. The PM is a semi-permeable structure composed of chitin and proteins. The Spodoptera exigua larval midgut cDNA expression library was constructed and screened with antiserum against S. exigua PM proteins. Two hundred and thirty-one positive clones were obtained. Sequence analysis showed that eight chitin binding protein and two invertebrate intestinal mucin were isolated from the Library. In this study,peritrophic membrane proteins were screened and structure analyzed for the development of new midgut targeting strategies for pests management.

关 键 词：甜菜夜蛾 CDNA表达文库 免疫学筛选 围食膜 序列分析 

分 类 号：Q78[生物学—分子生物学]