肠出血性大肠杆菌O157∶H7 LexA蛋白的表达、纯化及鉴定

沙建平; 邹全明; 张卫军; 杨珺; 毛旭虎; 郭刚; 罗萍; 刘开云; 陈洪章

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肠出血性大肠杆菌O157∶H7 LexA蛋白的表达、纯化及鉴定

论著 | 更新时间：2026-01-09

- 肠出血性大肠杆菌O157∶H7 LexA蛋白的表达、纯化及鉴定
- Expression, purification and identification of EHEC O157∶H7 LexA protein
- 陆军军医大学学报 2008年30卷第12期页码：1114-1117
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  中图分类号： R341;R378.21;R392-33
- 纸质出版：2008
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沙建平,邹全明,张卫军,杨珺,毛旭虎,郭刚,罗萍,刘开云,陈洪章.肠出血性大肠杆菌O157∶H7 LexA蛋白的表达、纯化及鉴定[J].陆军军医大学学报(原第三军医大学学报),2008,30(12):1114-1117.

　SHA Jian-ping,ZOU Quan-ming,ZHANG Wei-jun,YANG Jun,MAO Xu-hu,GUO Gang,LUO Ping,LIU Kai-yun,CHEN Hong-zhang.Expression, purification and identification of EHEC O157∶H7 LexA protein[J].J Amry Med Univ (J Third Mil Med Univ),2008,30(12):1114-1117.
沙建平,邹全明,张卫军,杨珺,毛旭虎,郭刚,罗萍,刘开云,陈洪章.肠出血性大肠杆菌O157∶H7 LexA蛋白的表达、纯化及鉴定[J].陆军军医大学学报(原第三军医大学学报),2008,30(12):1114-1117. DOI：

　SHA Jian-ping,ZOU Quan-ming,ZHANG Wei-jun,YANG Jun,MAO Xu-hu,GUO Gang,LUO Ping,LIU Kai-yun,CHEN Hong-zhang.Expression, purification and identification of EHEC O157∶H7 LexA protein[J].J Amry Med Univ (J Third Mil Med Univ),2008,30(12):1114-1117. DOI：

摘要

目的对EHEC O157∶H7 LexA进行表达、纯化，并检测其免疫活性。方法 lexA基因片段插入表达载体pET22b(+)，在E.coli BL21(DE3)中表达。包涵体经洗涤并用8 mol/L尿素溶解，Heparin Agrose亲合柱层析为第一步纯化，Superdex75凝胶过滤层析作为第二步精细纯化，HPLC测定LexA蛋白的浓度，将纯化的LexA蛋白经注射途径免疫家兔，制备兔抗lexA血清，采用免疫双扩、ELISA及Western blot分析LexA的免疫活性。结果 lexA以包涵体形式表达，表达产量高达总菌体蛋白的40%左右

经Heparin Agrose亲合柱层析和凝胶过滤层析两步组合纯化目的蛋白，经HPLC测定目的蛋白的最终纯度为98%，表达及纯化的lexA具有良好的免疫活性。结论在E.coli BL21(DE3)中表达的LexA蛋白具有良好的免疫活性。

Abstract

Objective To clone lexA gene from enterohemorrhagic Escherichia coli (EHEC) O157∶H7

and express and purify it

and measure its immunogenicity. Methods The objective fragment was amplified from standard EHEC O157∶H7 cells by PCR

and then linked with the vector pET22b(+). The objective protein was induced to express in the BL21(DE3) cells after transfection. The expressed protein was purified by 2 steps of Heparin Agrose chelate affinity chromatography and gel filtration chromatography respectively. The purified lexA protein was used to immune the rabbits to prepare the polyclonal antibody. ELISA and Western blotting was used to detect the obtained serum. Results The expressed protein was found in insoluble form

accounted for 40% of the total bacteria protein. The final purity was 98%

which was determined by HPLC. The obtained protein was confirmed to have sound immunogenicity. Conclusion Our obtained recombinant LexA protein expressed in E. coli BL21(DE3) has good immunological activity.

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