摘要
从230个具有ELISA阳性反应的细胞株获得了11株具有高效价的单克隆抗体, 其中2B4显示较强结合作用,并均能与分生孢子、芽管和附着胞表面结合。Western blotting分析表明,单抗2B4能与孢子芽管表面的1%SDS提取物中的15kDa的蛋白结合。免疫金定位发现该蛋白确是在病菌各个形态阶段广泛存在的分泌性蛋白。采用2B4从分生孢子cDNA表达文库中筛选获得6个阳性克隆。该克隆MP1表达的蛋白抗原与实际存在的15KDa是相一致的。其基因克隆及功能分析正在研究之中。
Eleven high effective monoclonal antibodies to Magnaporthe grisea were screened out from 230 hybrid cell strains showing positive reaction in ELISA tests, among which monoclonal antibody 2B4 showed more highly banding to cell wall surfaces of conidia, germ tubes, appressoria, germ tubes, appressoria or/and second infective hypha by indirect-immunofluorescence test. Ultrastructural analysis and the antigen localisition of monoclonal antibody 2B4 with immuno-gold labeling found this antigen protein exists extensively on cell walls of the differrent life stages above, and is a secretion protein. 6 positive cDNAs, MP1, MP2, MP3, MP4, MP5 and MP6 were screened out with 2B4 from conidium cDNA expression library. A 31kDa protein expressed from a positive clone MP1 in a prokaryotic in vitro translation system was consistent with a 15kda protein from the extractive of the fungal surfaces, and its gene cloning and functional analysis are undertaking.
出处
《菌物系统》
CSCD
北大核心
2002年第2期215-222,共8页
Mycosystema
基金
国家自然科学基金(30080021)项目资助
