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在枯草杆菌芽胞表面展示P74蛋白膜外肽段 被引量：1

Outer Membrane Peptide of P74 Displayed on the Surface of Bacillus subtilis Spore
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摘要 目的:对家蚕核型多角体病毒(BmNPV)囊膜蛋白P74膜外肽段与枯草芽胞杆菌(Bacillus subtilis)CotC与进行融合表达,制备表面展示有P74蛋白的重组芽孢,为深入研究该重组芽孢的功能提供基础。方法:将CotC基因与BmNPV P74膜外编码序列进行融合,构建表达CotC-P74融合蛋白的重组质粒pJS700-p74。通过双交换使该重组质粒中的CotC-P74表达盒整合到枯草芽胞杆菌染色体淀粉酶基因位点,并对发生同源重组后的菌株进行筛选和PCR鉴定。结果:PCR结果表明CotC-P74成功地整合到枯草芽孢杆菌基因组上,通过作者实验室制备的P74多抗对诱导后的重组芽孢衣壳总蛋白进行Western blot分析,结果在49 kDa位置处能杂交到一条特异的蛋白带。结论:P74蛋白胞外肽段成功地展示在枯草芽胞杆菌芽胞表面。 Objective： To display P74 on the surface of Bacillus subtilis （ B. subtilis ） spores for further functional analysis. Method： A recom- binant integrative plasmid named as pJS700 -p74,carrying cotC- p74 under the control of the cotC promoter, was constructed. Competent cells B. subtilis 168 （trp - ）were transformed with recombinant pJS700 -p74, which generated the resulting amylase inactivated mutant for a double crossover. Result：The disruption of amyE and the insertion of cotC -p74 in the amyE locus were confirmed by PCR, and a specif- ic hand about 49 kDa was detected in the recombinant spores extracts using P74 antibodies. Conclusion：P74 was displayed on B. subtilis spore surface successfully.
作者 李国辉 王鹏 唐琦 胡朝阳 黄国平
机构地区 江苏大学生命科学研究院
出处 《生物技术》 CAS CSCD 北大核心 2012年第5期25-29,共5页 Biotechnology
基金 国家自然科学基金项目(No.31000080) 江苏大学高级人才启动基金项目(No.09JDG057)资助
关键词 家蚕核型多角体病毒 P74 枯草芽孢杆菌 芽孢 CotC Bombyx mori nucleopolyhedrovirus P74 Bacillus subtilis spore CotC
分类号 Q812 [生物学—生物工程]
  • 相关文献

参考文献21

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共引文献1

同被引文献45

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生物技术
2012年 第5期

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