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目的 筛选和鉴定日本血吸虫特异性抗原基因。方法 采用日本血吸虫病流行区对再感染具有抵抗力的个体混合血清 ,对 Sjc DNA文库进行免疫学筛选 ,对获得的阳性克隆 c DNA插入片段进行 PCR鉴定和 PCR直接序列分析 ,通过互联网将获得的 EST序列送入 NCBI Gen Bank进行同源性检索 ,并将发现的未知基因 EST序列送入 NCBI db EST,以获得 EST编号和 Gen Bank进入编号。结果 通过对大约 2 .5× 10 4个噬菌体斑的免疫学筛选 ,获得了 5个阳性克隆 ,PCR直接序列分析表明 ,在这 5个阳性克隆中 ,1个为编码 Sj GST的基因克隆 ,另外 4个为 S.j未知抗原基因克隆 ,其 EST序列已被 Gen Bank接受 ,并获得进入编号。结论 应用对再感染具有抵抗力的个体混合血清 ,从 Sjc DNA文库中筛选到 4个日本血吸虫未知抗原基因重组克隆 ,为血吸虫疫苗候选抗原分子的研究提供了新的材料
Objective To screen and identify Schistosoma japonicum specific antigen gene. Methods Sjc DNA library was screened by immunization against resistant and susceptible individuals in the endemic area of Schistosoma japonicum. The positive clones were identified by PCR and sequenced directly by PCR. The obtained cDNA fragments The EST sequences are sent to NCBI Gen Bank for homology search, and the EST sequences of the unknown genes found are sent to NCBI db EST to obtain EST numbers and Gen Bank accession numbers. Results Five positive clones were obtained by immunological screening of about 2.5 × 10 4 phage plaques. PCR direct sequence analysis showed that among the five positive clones, one was a gene clone encoding Sj GST. In addition, 4 were Sj unknown antigen gene clones, and their EST sequences were accepted by Gen Bank and accession numbers were obtained. Conclusions Four serotypes of Schistosoma japonicum, which are unknown antigen genes, were screened from the Sjc DNA library using the mixed sera of restimulated individuals, which provided a new material for studying the candidate antigen molecules of schistosoma vaccine