日本血吸虫成虫抗原基因的获取及生物信息学分析

Acquisition and bioinformatics of analysis of gene encoding antigen for vaccine against adult S.japonicum

目的筛选新的日本血吸虫(Schistosoma japonicum,Sj)候选抗原基因。方法以Sj雄虫成虫DNA为模板,设计引物扩增并纯化特异性片段,构建pBS-T克隆载体,将其克隆入大肠杆菌DH5α菌株,筛选阳性克隆,初步进行菌落PCR鉴定后测序,与EBI数据库中的已知序列比对分析,并用蛋白质分析软件预测其结构与功能。结果利用日本血吸虫cDNA序列设计并合成引物,以Sj雄虫DNA为模板,成功地扩增和克隆出一个目的基因。通过菌落PCR检验和测序,BLAST比对分析显示其与日本血吸虫22.6kDa抗原分子具有很高同源性,氨基酸预测分析其具有潜在螺旋跨膜和酪氨酸激酶磷酸化位点。结论成功克隆一个目的基因,通过氨基酸预测和功能分析,预测其在血吸虫病免疫防治领域具有一定的意义。

Objective To screen a new effective gene of candidate vaccine against Schistosoma japonicum. Methods The template of male S.japonicum DNA and the designed primers were used to amplify the specific fragment, which was purified and cloned to the E. coli DH 5a. The inserts of the positive clones were identified firstly by PCR and further sequenced, and data were analyzed by internet Nueleotide BLAST software of EBI and Expert Protein Analysis System of Swiss Institute of Bioinformaties. Results A gene fragment was successfully cloned and the structure of nucleic acid and function was predicted. Conclusion Further study in screening and identifying some sehlstosomal genes encoding candidate antigens for vaccines against Schistosoma japonicum be conducted.