| 幽门螺杆菌Lpp20重组蛋白的表达、纯化及其免疫活性初步研究 |
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| 引用本文： | 靖吉芳,张艳,刘志杰,于文. 幽门螺杆菌Lpp20重组蛋白的表达、纯化及其免疫活性初步研究[J]. 免疫学杂志, 2010, 26(5) |
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| 作者姓名： | 靖吉芳 张艳 刘志杰 于文 |
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| 作者单位： | 南华大学病原生物学研究所,衡阳,421001;南华大学病原生物学研究所,衡阳,421001;南华大学病原生物学研究所,衡阳,421001;南华大学病原生物学研究所,衡阳,421001 |
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| 基金项目： | 湖南省自然科学基金优秀面上项目(06JJ2093);;湖南省教育厅优秀青年项目(08B067) |
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| 摘 要： | 目的表达幽门螺杆菌(Helicobacter pylori,H.pylori)脂蛋白Lpp20基因的重组蛋白,并检测其免疫活性。方法应用PCR技术从H.pylori标准株26695染色体DNA中扩增出Lpp20的编码基因片段,将其克隆至表达载体pGEX-6P-2上,在大肠杆菌(Escherichia coli,E.coli)BL21中表达并进行GST亲和层析纯化,用Western blot方法检测纯化产物重组Lpp20(recombinantLpp20,rLpp20)的免疫反应性。免疫C57BL/6小鼠后,以rLpp20为包被抗原建立间接ELISA法对免疫小鼠血清进行特异性抗体检测;ELISA双抗体夹心法检测小鼠脾淋巴细胞培养上清中IFN-γ、IL-2、IL-4水平;MTT比色法检测小鼠脾淋巴细胞增殖反应等指标以分析rLpp20的免疫活性。结果扩增的lpp20全长528 bp,与GenBank上公布的H.pylori 26695株lpp20序列作BLAST比较,结果完全一致;成功构建了pGEX-6P-2/Lpp20重组质粒,表达的rLpp20 M_r 43 000,纯化产物可被鼠抗H.pylori抗体识别;以...
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| 关 键 词： | 幽门螺杆茵 Lpp20 重组蛋白 蛋白表达 免疫活性 |
Expression and purification of the Lpp20 recombinant protein of Helicobacter pylori and primary study on its immunocompetence |
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| JING Jifang,ZHANG Yan,LIU Zhijie,YU Wen Institute of Pathogenic Biology,University of South China,Hengyang ,China. Expression and purification of the Lpp20 recombinant protein of Helicobacter pylori and primary study on its immunocompetence[J]. Immunological Journal, 2010, 26(5) |
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| Authors: | JING Jifang ZHANG Yan LIU Zhijie YU Wen Institute of Pathogenic Biology University of South China Hengyang China |
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| Affiliation: | JING Jifang,ZHANG Yan,LIU Zhijie,YU Wen Institute of Pathogenic Biology,University of South China,Hengyang 421001,China |
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| Abstract: | We aimed to express recombinant Lpp20 protein of Helicobacter pylori and study its immunocompetence.We amplified Lpp20 gene by polymerase chain reaction(PCR) and obtained Lpp20 gene fragments(528 bp).Then subcloned the fragment into the expression vector pGEX-6P-2 to generate recombinant plasmid pGEX-6P-2/Lpp20.Recombinant Lpp20(rLpp20) was expressed in Escherichia coli(E.coli) BL21 and purified by GST affinity chromatograph,thus we obtained a soluble fusion protein with molecular weight about 43 000.Then w... |
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| Keywords: | Helicobacter pylori Lpp20 Recombinant protein Protein expression Immunocompetence |
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