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目的寻找东方田鼠抗日本血吸虫抗性相关靶基因。方法利用东方田鼠血清以亲和淘洗方法筛选日本血吸虫成虫噬菌体展示cDNA文库,将所得阳性克隆进行测序及生物信息学分析,确定目的基因。结果经3轮筛选,特异性噬菌体得到有效富集(857倍),随机挑取58个克隆进行序列测定,获得了10条表达序列标签(EST)序列,其中7条与GenBank中登陆的日本血吸虫基因序列具有99%～100%同源性,1条与类人猿锌指蛋白基因有82%同源性。功能预测结果表明,大部分EST编码蛋白或多肽的功能主要是参与血吸虫基因表达调控。结论发现了多个可能与东方田鼠抗血吸虫抗性相关的靶基因,为进一步阐述东方田鼠抗血吸虫病的分子机理奠定了基础。
Objective To search for the target gene of Microtus fortis against Schistosoma japonicum resistance. Methods The phage display cDNA library of adult Schistosoma japonicum was screened by affinity elutriation using the voles of Microtus fortis. The positive clones were sequenced and analyzed by bioinformatics to determine the target gene. Results After 3 rounds of screening, the specific phage was enriched (857-fold), 58 clones were randomly selected and sequenced. Ten ESTs were obtained, of which 7 were identical to those of S. japonicum landed on GenBank The gene sequence has 99% to 100% homology and one gene has 82% homology with the ape zinc finger protein gene. Functional prediction results show that most of the function of EST-encoded protein or polypeptide is involved in the regulation of gene expression in schistosomiasis. Conclusion A number of target genes related to resistance to T. orientalis were found, which laid the foundation for further elucidating the molecular mechanism of S. japonicus anti-schistosomiasis.