现代检验医学杂志2013，Vol.28Issue(1)：7-11,5.DOI:10.3969/j.issn.1671-7414.2013.01.003
幽门螺杆菌过氧化氢酶的重组表达、纯化及免疫特性研究
Recombinant Expression and Immunological Characteristcs of Helicobacter Pylori Catalase
屈玲 1黄萍 2苏明权 1郑善銮 1马越云 1郝晓柯1
作者信息
- 1. 第四军医大学西京医院全军临床检验医学中心,西安,710032
- 2. 第四军医大学口腔医院检验科,西安,710032
- 折叠
摘要
Abstract
Objective To prepare recombinant catalase protein (rKatA) of Helicobacter pylori (H. Pylori) expressed in E.
Coli,and investigate its immunological characteristics. Methods H. Pylori catalase gene was amplified by PCR and construe ted into pET 28a vector. rKatA was expressed in E coli BL21 (DE3) and purified by inclusion body washing and affinity chromatography (chelating sepharose fast flow). The immuno logical characteristics of rKatA was investigate by Western blot and ELISA. Results The catalase gene of H. Pylori cloned by PCR contained 1 518 base pairs molecule encoding 505 a mino acids. rKatA was expressed as inclusion body in E. Coli BL21 with about 20% expression rate and 58 000 molecular weight. The purity of rKatA was up to 85% after affinity chromatography. Recombinant protein was specifically identified by rabbit antiserum against H. Pylori, and high titer anti rKatA antibody came from immunized rabbit with rKatA could recognized various H. Pylori strains. Conclusion rKatA was successfully prepared and maintained good immunological characteristics,which would be useful for further research on pathogenic mechanism, serodiagnosis and subunit vaccine of H. Pylori关键词
幽门螺杆菌/过氧化氢酶/重组表达/免疫学特性Key words
helicobacter Pylori/ catalase/ recombinant expression/ immunological characteristics分类
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屈玲,黄萍,苏明权,郑善銮,马越云,郝晓柯..幽门螺杆菌过氧化氢酶的重组表达、纯化及免疫特性研究[J].现代检验医学杂志,2013,28(1):7-11,5.
