| 甜菜夜蛾围食膜几丁质结合蛋白SeCBP66在不同体系中的表达 |
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| 引用本文： | 刘彬,李瑞军,郭巍,孙伟明,赵丹,单鸣,徐大庆.甜菜夜蛾围食膜几丁质结合蛋白SeCBP66在不同体系中的表达[J].河北农业大学学报,2012(4):80-84. |
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| 作者姓名： | 刘彬 李瑞军 郭巍 孙伟明 赵丹 单鸣 徐大庆 |
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| 作者单位： | 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;河北农业大学生命科学学院;国家北方山区农业工程技术研究中心;青岛农业大学中美无脊椎动物细胞工程中心 |
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| 基金项目： | 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) |
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| 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。
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| 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 |
Expression of peritrophic membrane chitin binding protein SeCBP66 of Spodoptera exigua in different systems |
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| LIU Bin,LI Rui-jun,GUO Wei,SUN Wei-ming,ZHAO Dan,SHAN Ming,XU Da-qing.Expression of peritrophic membrane chitin binding protein SeCBP66 of Spodoptera exigua in different systems[J].Journal of Agricultural University of Hebei,2012(4):80-84. |
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| Authors: | LIU Bin LI Rui-jun GUO Wei SUN Wei-ming ZHAO Dan SHAN Ming XU Da-qing |
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| Institution: | 1.College of Plant Protection,Agricultural University of Hebei,Biological Control Center of Plant Diseases and Plant Pests of Hebei Province,Baoding 071001,China;2.College of Life Sciences,Agricultural University of Hebei, Baoding 071001,China;3.National Engineering Research Center for Agriculture in Northern Mountainous Areas, Baoding 071001,China;4.Center for Advanced Invertebrate Cell Engineering Collaborated Between Boyce Thompson Institute and Qingdao Agricultural University,Qingdao Agricultural University(QAU),Qingdao 266109,China) |
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| Abstract: | 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.pastoris GS115,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(HighFive).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. |
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| Keywords: | Spodoptera exigua SeCBP66 prokaryotic expression system P pastoris expression system insect cell lines |
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