蟑螂变应原基因的免疫学筛选及序列分析

Immunology screen and sequence analysis of cockroach allergen genes

目的 探讨从美洲蟑螂cDNA表达文库中获得的变应原基因对过敏性疾病的治疗价值。方法 用生物素 链霉亲合素系统 ,使用蟑螂过敏病人血清IgG4,对已构建的美洲蟑螂cDNA表达文库进行免疫学筛选 ,并对阳性克隆进行序列测定及同源性分析。结果 从美洲蟑螂cDNA文库筛选出 3个阳性克隆 ,同源性分析显示 ,3号克隆核苷酸序列及推导的氨基酸序列与美洲蟑螂主要变应原CrPI高度同源 ,一致性达 99% ,1、2号克隆与已知变应原无显著同源性 (score <2 0 0 ) ,提示其为新的蟑螂变应原基因 ,其中 1号为编码核糖体蛋白S12 ,2号为编码Rab11蛋白。结论 3号为克隆编码美洲蟑螂主要变应原CrPI,另外 2个克隆为新的蟑螂变应原基因 ,对这些基因的进一步亚克隆表达 。

Objective To obtain allergen genes from American coc kroach cDNA expression library. Methods Using Biotin streptavidin system, Cockroach cDNA library was screened with cockroach allergic patients′ IgG4, The positive clones were sequenced and analyzed.Results Three positive clones were obtained from cockroach cDNA library, DNA sequence analysis showed the nucleic acid sequence and putative amino acid sequence of third clone were highly homology with American cockroach major allergen Cr PI, identities was 99%, but no significant homology was found between the other two and all the know allergen sequences(score<200), the first one coded for ribosomal protein S12, the second one coded for Rab11.Conclusions The third clone coded for American cockroach major allergen Cr PI, The other two were new cockroach allergen genes. The subcloning of these genes will promote the diagnosis and therapy of allergic disease by recombinant allergen.