中国免疫学杂志2012，Vol.28Issue(5)：444-448,5.DOI:10.3969/j.issn.1000-484X.2012.05.015
日本柳杉花粉变应原CJP-6的重组表达和免疫反应性鉴定
Expression and immunoreactivity of Cryptomeria japonica pollen allergen, CJP-6
摘要
Abstract
Objective:To construct a recombinant prokaryotic expression vector to express Japanese cedar (Cryptomeria japoni-ca) pollen allergen, CJP-6, and to identify IgE-binding capacity of the recombinant allergen. Methods:The total RNA was extracted from C. Japonica pollen, and then the CJP-6 gene fragments were amplified by RT-PCR and cloned into vector pET-19b. The recombinant plasmids were transformed to E. Coli BL21 Star (DE3) pLysS. After induced by IPTG, the recombinant protein was purified through Ni2+ affinity chromatography. The IgE binding activity of rCJP-6 is assessed using sera from patients allergic to C. Japonica pollen, mite and Artemisia pollen by Dot blot and ELISA. Results:The recombinant allergen was expressed in E. Coli and purified through Ni2+ affinity chromatography successfully. The rCJP-6 has high affinity with IgE existing in sera of patients allergic to C. Japonica pollen, mite and Artemisia pollen. Conclusion;The rCJP-6, which has high IgE-binding activity, was expressed and purified. It will be a practical tool for clinical detection, immunotherapy and further study for allergic diseases.关键词
日本柳杉/CJP-6/变应原/交叉反应性/IgEKey words
Cryptomeria japonica/CJP-6 / Allergen/Cross-reactivity/ IgE分类
医药卫生引用本文复制引用
蒋建国,刘瑞玲,付永锋,程训佳..日本柳杉花粉变应原CJP-6的重组表达和免疫反应性鉴定[J].中国免疫学杂志,2012,28(5):444-448,5.基金项目
本文受国家高技术研究发展计划项目(2007AA02Z472)和卫生行业科研专项项目(20082001)资助 （2007AA02Z472）
