日本血吸虫尾蚴66～68kD单特异性血清对尾蚴cDNA文库的免疫学筛选

Immuno-screening of Schistosoma japonicum cercariae cDNA library by the sera of anti-soluble cercariae 66～68 kD antigens

目的:获得日本血吸虫(Schistosoma japonicum,Sj)尾蚴66～68 kD抗原的编码基因,为日本血吸虫疫苗和诊断研究提供材料。方法:以Sj尾蚴66～68 kD抗原免疫家兔获得的单特异性血清为探针,对Sj尾蚴cDNA文库进行免疫筛选,将所获阳性克隆的插入片断进行PCR扩增、琼脂糖电泳初步鉴定,并挑选出4个强阳性克隆进行测序,通过互联网NCBI/BLAST进行核苷酸序列分析。结果:共获得21个持续阳性克隆,其中10个阳性克隆初步鉴定显示为单一扩增条带,且插入的cDNA片段大小分布于0.5～3.0 kb之间;4个强阳性克隆分别与日本血吸虫SJCHGC05187,SJCHGC05173,SJCHGC06989,SJCHGC01894显著同源。结论:获得了4种日本血吸虫相关的编码基因。

Objective To obtain the coding genes related to Schistosoma japonicum （Sj） cercariae 66 -68 kD antigens, and to provide antigens for diagnosis and vaccine of schistosomiasis. Methods Sj cercariae cDNA library was screened using the monospecific anti-sera of rabbit agaist soluble cercariae 66 -68 kD antigens as probes. The inserted cDNA fragments of the positive clones were amplified with PCR and identified by agarose gel electrophoresis. Four strong positive clones were further sequenced and analyzed through the internet NCBI/BLAST software. Results Twenty-one positive clones were obtained, 10 of which revealed a single band （0. 5 - 3.0 kb ）. The 4 strong positive clones showed high identity to SJCHGC05187, SJCHGC05173, SJCHGC06989, and SJCH- GC01894 at the nueleotide level. Conclusion Four coding genes related with Sj antigens are obtained.