黑胸大蠊与美洲大蠊交叉抗原成分比较

Comparison of Cross-reactive Antigenic Components in Periplaneta Fuliginosa and Periplaneta Americana

对黑胸大蠊与美洲大蠊体部浸出液交叉抗原成分进行分析 ,为利用有关美洲大蠊分子克隆的资料进行黑胸大蠊主要变应原的克隆、测序和重组表达做前期工作。制备纯种黑胸大蠊及美洲大蠊的体部浸出液 ,采用SDS PAGE (12 %分离胶 )分离蛋白成分 ,用Coomassi亮蓝染色 ,比较两种来源蛋白成分的差异。然后将蛋白电转至PVDF膜 ,以兔抗黑胸大蠊体部浸出液 (WBE )抗体作为第一抗体 ,进行Western免疫印迹检测 ,以判断两种蟑螂的交叉抗原成分。两种蟑螂体部浸出液蛋白成分的SDS PAGE带型略有差异 ,主要是条带密度不一致 ,而从其疏密分布 (分布型式 )上看 ,两者间存在相当多的分子量相同或相近组分。经SDS PAGE分离的条带多达近 30条 ,分子量相同的组分多达 2 0余条。免疫印迹表明 ,针对兔抗黑胸大蠊体部浸出液 (WBE )抗体 (IgG )有交叉反应的抗原成分多达 10几条。黑胸大蠊与美洲大蠊体部浸出液的蛋白成分或变应原组分间存在交叉反应成分。提示 ,利用有关美洲大蠊分子克隆的资料进一步对黑胸大蠊主要变应原进行克隆、测序或重组表达等分子生物学研究具备一定的可行性。

To determine the capability of molecular cloning,sequencing and recombinant expression of Periplaneta fuli- ginosa through a convenient way using the already available information of Periplaneta americana,we have carried out the assay of cross-reactive antigenic components in Periplaneta fuliginosa and Periplaneta americana Extracts of two kinds of cockroaches were prepared for SDS-PAGE and Western blot 12% separating gel and rabbit anti-Per. fuliginosa antibody as the first antibody were used respectively in SDS-PAGE and Western blot Band patterns of SDS-PAGE and Western blot were compared with each other to find any significant differences Little difference was found in the band patterns of two species of cockroaches with only a slightly different appearance in their densities More than 20 bands were identical on the SDS-PAGE gel,and more than 10 bands in the same molecular weight were seen on immunoblotted (Western blot) PVDF film Cross-reactive antigenic components do exist in two sub-species of cockroaches So,it is capable of carrying out the molecular cloning of Periplaneta fuliginosa using the molecular cloning information of Periplaneta americana