| 淋球菌porB和大肠杆菌ltB融合基因的构建、表达及其免疫活性 |
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| 引用本文： | 戴志兵,胡四海,陈敏,陆春雪,王玉峰,刘清南,张愉快,余敏君,朱翠明,李忠玉.淋球菌porB和大肠杆菌ltB融合基因的构建、表达及其免疫活性[J].微生物学报,2010,50(4):517-523. |
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| 作者姓名： | 戴志兵 胡四海 陈敏 陆春雪 王玉峰 刘清南 张愉快 余敏君 朱翠明 李忠玉 |
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| 作者单位： | 1. 南华大学,病原生物学研究所,衡阳421001
2. 南华大学,心血管疾病研究所,衡阳421001 |
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| 基金项目： | 国家自然科学基金（30771931）；湖南省教育厅科研基金（08C745） |
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| 摘 要： | 【目的】构建融合基因原核表达载体pET-30a/ltB-porB,并表达重组融合蛋白LTB-PorB,鼻饲途径免疫雌性BALB/c小鼠,分析重组融合蛋白的免疫活性,为研制抗淋病蛋白疫苗提供实验依据。【方法】构建大肠杆菌不耐热肠毒素B亚单位(LTB)与淋球菌外膜孔蛋白B(PorB)融合基因及LTB、PorB单基因pET-30a原核表达载体,在大肠杆菌BL21中表达重组蛋白;鼻饲途径免疫雌性BALB/c小鼠,检测体液免疫和细胞免疫水平。【结果】在大肠杆菌BL21中获得高效表达的重组蛋白;经鼻饲免疫小鼠后,重组融合蛋白LTB-PorB组生殖道黏膜产生的PorB特异性sIgA水平随免疫时间呈上升趋势,第42天A450值达0.66,明显高于对照组(P0.01),效价高达1∶1280;血清中产生的PorB特异性IgG第28天达最高,A450值为0.60,明显高于LTB和蛋白溶解液(Solution Buffer)对照组(P0.01),效价高达1:2560,但与PorB对照组血清IgG水平(A450:0.57)无明显差异(P0.05)。LTB-PorB组脾淋巴细胞刺激指数明显高于LTB和SolutionBuffer对照组(P0.05),但脾淋巴细胞诱生的IFN-γ水平与对照组无明显差异(P0.05)。【结论】重组融合蛋白LTB-PorB通过鼻饲途径免疫雌性BALB/c小鼠后,能诱导产生高水平的体液免疫和一定水平的细胞免疫。首次证实黏膜佐剂LTB可辅佐PorB诱导小鼠产生高水平的生殖道粘膜免疫。
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| 关 键 词： | 关键词：淋病奈瑟菌 孔蛋白B 大肠杆菌不耐热肠毒素B亚单位 基因融合 原核表达 免疫活性 |
| 收稿时间： | 2009/12/6 0:00:00 |
| 修稿时间： | 2009/12/31 0:00:00 |
Construction, expression of Neisseria gonorrhoeae porB and Escherichia coli ltB fusion gene and analysis of its Immunocompetence |
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| Zhibing Dai,Sihai Hu,Min Chen,Chunxue Lu,Yufeng Wang,Qingnan Liu,Yukuai Zhang,Minjun Yu,Cuiming Zhu and Zhongyu Li.Construction, expression of Neisseria gonorrhoeae porB and Escherichia coli ltB fusion gene and analysis of its Immunocompetence[J].Acta Microbiologica Sinica,2010,50(4):517-523. |
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| Authors: | Zhibing Dai Sihai Hu Min Chen Chunxue Lu Yufeng Wang Qingnan Liu Yukuai Zhang Minjun Yu Cuiming Zhu and Zhongyu Li |
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| Institution: | Pathogenic Biology Institute, University of South China, hengyang, 421001, China;Pathogenic Biology Institute, University of South China, hengyang, 421001, China;Pathogenic Biology Institute, University of South China, hengyang, 421001, China;Pathogenic Biology Institute, University of South China, hengyang, 421001, China;Pathogenic Biology Institute, University of South China, hengyang, 421001, China;Pathogenic Biology Institute, University of South China, hengyang, 421001, China;Pathogenic Biology Institute, University of South China, hengyang, 421001, China;Pathogenic Biology Institute, University of South China, hengyang, 421001, China;Pathogenic Biology Institute, University of South China, hengyang, 421001, China;Pathogenic Biology Institute, University of South China, hengyang, 421001, China |
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| Abstract: | Objective To construct prokaryotic fusion gene expression vector pET-30a /ltB-porB,express the recombinant fusion protein LTB-PorB and analyze the immunocompetence of the recombinant fusion protein in female BALB /c mice through intranasally immunization.MethodsB subunit of Escherichia coli heat-labile enterotoxin(LTB) and Neisseria gonorrhoeae Porin B(PorB) fusion gene,LTB gene and PorB gene were cloned into prokaryotic vector pET-30a.The recombinants were identified by Polymerase Chain Reaction(PCR),enzym... |
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| Keywords: | Keywords: Neisseria gonorrhoeae Porin B B subunit of Escherichia coli heat-labile enterotoxin gene fusion prokaryotic expression Immunocompetence |
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