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Expression, purification and characterization of the American cockroach Cr PI allergen
Author(s): 
Pages: 13-17
Year: Issue:  1
Journal: ACTA ENTOMOLOGICA SINICA

Keyword:  美洲大蠊重组变应原Cr PI蛋白表达亲和层析免疫印迹;
Abstract: 以阳性噬菌体克隆为模板,通过PCR扩增出目的基因片段并克隆入T载体,经测序证实为美洲大蠊Periplaneta americana变应原Cr PI后,将该基因亚克隆入表达载体pGEX-5X-1.美洲大蠊变应原Cr PI在大肠杆菌中得到高效表达,但主要以包涵体形式存在于沉淀中.目的蛋白溶于6 mol/L盐酸胍并经稀释复性后,经Glutathione SepharoseTM 4B亲和层析,纯度达90%以上.以蟑螂过敏病人血清进行免疫印迹检测,结果显示重组变应原具有良好的IgE结合活性.
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