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目的 通过构建日本血吸虫成虫 c DNA表达文库 ,免疫筛选全长目的基因 ,研究其结构和功能。 方法 采用 TRIZOL试剂提取日本血吸虫成虫总 RNA,Oligotex m RNA Midi Kit分离 m RNA,并用 Stratagene的建库试剂盒构建日本血吸虫 c DNA表达文库。 结果 构建的日本血吸虫成虫 c DNA表达文库容量为 2× 10 6 pfu/ ml,扩增后文库的滴度为 1.4× 10 7pfu/ ml。文库的重组率达到 96%以上 ,插入片段平均长度为 1.4kb。 结论 本文库可望用于日本血吸虫疫苗候选基因及诊断抗原分子的筛选
Objective To construct the cDNA library of Schistosoma japonicum adult c worm and screen the full-length target gene by immunoprecipitation to study its structure and function. Methods Total RNA of adult Schistosoma japonicum was extracted with TRIZOL reagent, m RNA was extracted with Oligotex m RNA Midi Kit, and constructed with Stratagene library. Results The constructed cDNA library of Schistosoma japonicum c DNA expression was 2 × 10 6 pfu / ml, and the amplified library titer was 1.4 × 10 7 pfu / ml. The recombination rate of the library reached more than 96%, and the average length of the insert was 1.4kb. Conclusion This library is expected to be used in the screening of Schistosoma japonicum vaccine candidate genes and diagnostic antigen molecules