甜菜夜蛾围食膜几丁质结合蛋白SeCBP66在不同体系中的表达  

Expression of peritrophic membrane chitin binding protein SeCBP66 of Spodoptera exigua in different systems

作　　者：刘彬[1] 李瑞军[1] 郭巍[1,2,3] 孙伟明[1] 赵丹[1] 单鸣[4] 徐大庆[2] 

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071000 [3]国家北方山区农业工程技术研究中心,河北保定071001 [4]青岛农业大学,中美无脊椎动物细胞工程中心,山东青岛266109

出　　处：《河北农业大学学报》2012年第4期80-84,共5页Journal of Hebei Agricultural University

基　　金：国家现代农业(花生)产业技术体系项目;“973计划项目”农业生防微生物制剂的合成与作用机理及定向改造(2009CB118902)

摘　　要：分析甜菜夜蛾(Spodoptera exigua)围食膜几丁质结合蛋白SeCBP66氨基酸序列,利用PCR技术扩增获得缺失信号肽编码区围食膜几丁质结合蛋白的基因△(s)-secbp66。将△(s)-secbp66克隆到表达载体pET28a和pPIC9K,分别转化大肠杆菌BL21和毕赤酵母GS115进行原核及真核表达;将△(s)-secbp66克隆到质粒pFastBacTM,转化大肠杆菌DH10Bac构建重组Bacmid,转染昆虫细胞系Sf9及BTI-TN-5B1-4(HighFive)从而在昆虫细胞系中进行表达。结果表明:SeCBP66在原核细胞中未见表达,在酵母细胞中表达产物呈弥散状,而在昆虫细胞系中则能稳定的表达116kDa的特异蛋白。Amino acid sequence of chitin-binding protein SeCBP66 of Spodoptera exigua peritrophic membrane was analyzed and a △（s）-secbp66 encoding SeCBP66 without signal peptide was get by PCR method from the midgut cDNA library of Spodoptera exigua. The △（s）- secbp66 gene was cloned into pET28a and pPIC9K and transformed into E. coli BL21 ,P. [asto- ris GSllS,resulting recombinant E. coli and P. pastoris,respectively. Using Bac to Bac system, the △（s）-secbp66 gene was transfected into insect cell lines Sf9 and BTI-TN-5B1-4（High- Five）. The results showed SeCBP66 was expressed in P. pastoris and insect cell lines, but not in E. coli. However, SeCBP66 appeared as a dispersion of unequal size in P. pastoris while a stable expression was showed as a molecular weight 116 kDa in insect cell lines.

关 键 词：甜菜夜蛾 SeCBP66蛋白 原核表达系统 毕赤酵母表达系统 昆虫细胞系 

分 类 号：Q786[生物学—分子生物学]