东方田鼠抗日本血吸虫抗性相关靶基因筛选  

Screening the target genes of Schistosoma japonicum related to the naturely resistance of Microtus fortis

作　　者：孙毅[1] 孙焕[1] 贾人初[1] 刘金明[1] 苑纯秀[1] 石耀军[1] 陆珂[1] 李浩[1] 金亚美[1] 林矫矫[1] 

机构地区：[1]中国农业科学院上海兽医研究所、国家防治动物血吸虫病专业实验室、农业部动物寄生虫学重点开放实验室,上海200232

出　　处：《中国血吸虫病防治杂志》2008年第1期26-31,共6页Chinese Journal of Schistosomiasis Control

基　　金：上海市科学技术委员会科研计划项目(054909002);科技部自然资源平台项目(2005DKA21104)

摘　　要：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 look for the genes of Schistosoma japonicum related to the Schistosomaresistance of Microtus fortis. Methods The fresh sera of Microtus fortis were used to screen a T7 phage display eDNA library from worms of Schistosoma japonicum established in our lab. The positive clones were sequenced and functionally analysed through bioinformatics. Results The specific phages binding to the sera of Microtus fortis were enriched 857-fold after three rounds of biopanning, and 58 positive clones picked at random were sequenced and 10 ESTs were obtained. BLASTn results showed that 7 ESTs had 99 %-100% similarity to the genes of Shistosoma japonicurn reported in Genl3ank and 1 EST had 82% similarity to a zinc finger protein encoden gene from Pan troglodytes. The results of these ESTs function prediction indicated most of them were involved in the regulation of gene expresion of Schistosoma ja ponicum. Conclusions Several target genes of Schistosoma ja ponicum related to the Schistosoma-resistance of Microtus fortis are obtained and those would lay foundation to expatiate the native resistance meehnism of Microtus fortis to Schistosoma ja ponicum.

关 键 词：日本血吸虫 东方田鼠 噬菌体展示cDNA文库 靶基因 亲和淘洗 

分 类 号：R383.24[医药卫生—医学寄生虫学]