Preparation of recombinant CagL protein of Helicobacter pylori and its neutralization antibody

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Preparation of recombinant CagL protein of Helicobacter pylori and its neutralization antibody

更新时间：2026-01-09

- Preparation of recombinant CagL protein of Helicobacter pylori and its neutralization antibody
- Journal of Army Medical University Vol. 31, Issue 2, Pages: 109-112(2009)
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  CLC： R378;R392-33;R392.1
- Published：2009
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钟情,邹全明,郭刚,章金勇,张晓丽.重组表达幽门螺杆菌致病岛CagL蛋白及其中和抗体初步研究[J].陆军军医大学学报(原第三军医大学学报),2009,31(02):109-112.

　ZHONG Qing,ZOU Quan-ming,GUO Gang,ZHANG Jin-yong,ZHANG Xiao-li.Preparation of recombinant CagL protein of Helicobacter pylori and its neutralization antibody[J].J Amry Med Univ (J Third Mil Med Univ),2009,31(02):109-112.
钟情,邹全明,郭刚,章金勇,张晓丽.重组表达幽门螺杆菌致病岛CagL蛋白及其中和抗体初步研究[J].陆军军医大学学报(原第三军医大学学报),2009,31(02):109-112. DOI：

　ZHONG Qing,ZOU Quan-ming,GUO Gang,ZHANG Jin-yong,ZHANG Xiao-li.Preparation of recombinant CagL protein of Helicobacter pylori and its neutralization antibody[J].J Amry Med Univ (J Third Mil Med Univ),2009,31(02):109-112. DOI：

摘要

目的重组表达幽门螺杆菌（Helicobacter pylori，Hp）cagL基因，并制备兔抗多克隆抗体及阻断Hp黏附宿主细胞实验。方法用PCR方法扩增出cagL基因片段，将目的基因构建至表达载体pET22b中并诱导表达。采用亲和层析法纯化蛋白，以纯化蛋白为抗原免疫家兔，制备兔抗CagL多克隆抗体血清，并用多克隆抗体血清进行阻断Hp黏附宿主细胞实验及细胞因子IL-8活性分析。结果成功构建表达纯化了CagL重组蛋白，制备CagL蛋白兔抗多克隆血清，并初步验证了多克隆抗体血清在体外和Hp黏附作用的关系。与对照菌相比，重组菌CagL在25×10

3

附近均出现新的蛋白表达条带，且诱导6 h时表达量最高。UVP扫描分析目的蛋白占菌体总蛋白的35％～40％。SDS-PAGE显示重组蛋白CagL主要以包涵体形式存在于超声沉淀中。ELISA结果显示兔抗CagL多克隆抗体的滴度为1∶16 000 EU。Giemsa染色后显微镜观察可见Hp菌与HeLa细胞呈典型的聚集性黏附，抗体中和后Hp与HeLa细胞未见黏附，加入DH5α的HeLa细胞也未见细菌黏附。结论多克隆抗体血清对Hp黏附定植有一定阻断作用，并且可以减少Hp促进细胞分泌IL-8的能力。

Abstract

Objective To study the function of CagL protein of recombinant Helicobacter pylori (Hp) through preparing rabbit anti-CagL specific polyclonal antibody and blocking the adhesion of Hp with host cells. Methods CagL gene was amplified by PCR

then constructed into prokaryotic expression vector pET-22b. Expressed protein of CagL was purified by affinity chromatography

and then used to immunize rabbits to prepare anti-CagL polyclonal antibody serum

so as to detect whether CagL polyclonal antibody serum can block the adhesion of Hp with host cells and analyze the activity of IL-8. Results Recombinant CagL protein was successfully constructed and purified. CagL polyclonal antibody serum played a stronger role in blocking the adhesion of Hp with host cells

and can induce host cells to secrete IL-8. Conclusion Our study suggests that CagL plays a role in Hp pathogenicity and colonization

and secretion of IL-8.

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