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目的 :分离 Sj2 2 .6抗原基因 ,构建 Sj2 2 .6表达克隆。方法 :抗体筛选 Sj成虫 c DNA库 ,PCR扩增目的基因片段 ,亚克隆入表达载体 p GEX- 1λt。对重组体进行诱导表达并对表达产物进行 SDS- PAGE和 Western blot分析。结果 :PCR扩增产物为约 930 bp的 DNA条带。亚克隆入p GEX- 1λt,获得两个表达克隆 p GSj6和 p GSj2 4 ,特异性表达产物为 2 2 .6k Da抗原。重组体的表达方式为分离表达。结论 :从日本血吸虫成虫 c DNA库筛选到 Sj2 2 .6k Da克隆化基因 ,并构建了表达克隆 p GSj2 4和 p GSj6。
Objective: To isolate Sj2 2 .6 antigen gene and construct Sj2 2 .6 expression clone. Methods: Antibody was used to screen the cDNA library of Sj adult c DNA. The target gene fragment was amplified by PCR and subcloned into the expression vector pGEX-1λt. The recombinant was induced to express and the expressed product was analyzed by SDS-PAGE and Western blot. Results: The PCR product was about 930 bp DNA band. Subcloned into pGEX-1λt to obtain two expression clones p GSj6 and p GSj2 4, the specific expression product is 226k Da antigen. Recombinant expression is expressed separately. CONCLUSION: The cloned Sj2-2.6 kDa gene was screened from the c DNA library of adult Schistosoma japonicum and the expression clones p GSj2 4 and p GSj6 were constructed.