中国免疫学杂志2011，Vol.27Issue(8)：726-730,5.DOI:10.3969/j.issn.1000-484X.2011.08.011
欧蓍草花粉主要过敏原Par j1的重组表达及鉴定
Cloning and expression of the Parietaria judaica pollen allergen Par j 1
摘要
Abstract
Objective:To express and purify the Parietaria judaica pollen major allergen Par j1.Methods:We obtained the nucleotide and amino acid sequence according to the published allergen number(O04404)in the GenBank.Codons optimized by DNA star software.The optimized gene was synthesized and cloned into the expression vector pET-44a and transformed into E.coliRosetta strain.Protein expression parameters were optimized on inducer concentration of IPTG,expression temperature and induction time.Recombinant protein was purified by affinfity chromatography and identified by Western blot.Results:The recombinant plasmid pET-44A=/Par j1.0102 was successfully constructed whichproved by PCR amplification and sequencing.Par j1.0102 protein was recombinantly expressed with StrepⅡ tag under the best situation.Western blot showed recombinant protein could specifically bind to the StrepⅡ tag antibody.Conclusion:The Par j1.0102 protein is correctly expressed and the expressed protein could be used for diagnosis and immunotherapy of Parietaria judaica allergic patients with its allergenicity attenuated.关键词
欧蓍草/花粉过敏原/Par j 1.0102/密码子优化/克隆/表达/纯化Key words
Parietaria judaica/Pollen allergen/Par j1.0102/Codon optimization/Clone/Expression/Purification分类
医药卫生引用本文复制引用
林德球,程江丽,邹泽红,何颖,陶爱林..欧蓍草花粉主要过敏原Par j1的重组表达及鉴定[J].中国免疫学杂志,2011,27(8):726-730,5.基金项目
本文为国家科技重大专项重大课题(2008ZX08011-005)及重点课题(2009ZX08011-004B)、国家自然科学基金项目(30771240)和广州市教育系统科研创新学术团队(B94118)课题 （2008ZX08011-005）
