卫氏并殖吸虫成虫cDNA文库的单抗筛选

Immunoscreening of Paragonimus westermani adult cDNA library using monoclone antibodies

目的 从卫氏并殖吸虫成虫cDNA文库中筛选并鉴定出可用于免疫诊断和免疫预防的基因克隆。方法 采用预吸收的抗肺吸虫成虫单克隆抗体筛选多次后得到 5个阳性克隆 ,经PCR扩增后测定其插入片段的大小。用辅助噬菌体做体内剪切 ,以抗生素平板筛选含重组质粒的阳性菌落。其中 3个克隆测定其DNA序列 ,用BLAST软件对所得DNA序列进行同源性比较。结果 得到 1,2 ,4三个不同阳性克隆 ,其长度分别为 1783bp、397bp和 1132bp ,分别与卫氏并殖吸虫卵黄铁蛋白 (P wyolkferritin)基因、鞘脂激活蛋白A(DictyosterliumdiscoideumsaposinA)基因和曼氏血吸虫主要卵抗原 (Smmajoreggantigen -P4 0 )基因同源。 结论 本研究首次报道用抗肺吸虫成虫单克隆抗体筛选卫氏并殖吸虫成虫cDNA文库 ,所获克隆1、2。

Aim To creen and identify gene clones from the cDNA library of the adult of Paragonimus westermani(PwA) for immunodiagnosis and immunoprophylaxis Methods PwA cDNA library was screened with the monoclone antibodies(McAB) pre absobed by the extract E coli XL1 Blue Positive clones were amplified with PCR,the PCR products were mesured by eletctrophoresis and excised automatically in the help phages Three obtained positive clones named clone 1,2,4 were sequenced and analysed with BLAST software Result Clone 1,2,4 were obtained,the sizes of their insert fragments were about 1700,400,1000bp They were homologous with P w yolk ferritin gene,saposin A gene and Sm p40 gene respectively Conclusion The screen of PwA cDNA library with monoclone antibodies(McAB) pre absobed has been firstly reported The clone 1,2,4 that we obtained may be appled to immunodiagnosis and immunoprophylaxis of Paragonimiasis