东方田鼠肝脏裂解物筛选日本血吸虫成虫噬菌体展示cDNA文库  

Screening a phage display cDNA library of Schistosoma japonicum with extract from liver of Microtus fortis

作　　者：孙焕[1] 刘金明[1] 孙帅[1] 宋震宇[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：《中国预防兽医学报》2008年第8期592-596,617,共6页Chinese Journal of Preventive Veterinary Medicine

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：东方田鼠对血吸虫具有天然抗性。为寻找东方田鼠抗血吸虫抗性相关靶基因,探索其分子机理,我们将日本血吸虫童虫与东方田鼠不同组织/器官裂解物共培养,比较它们对血吸虫童虫的杀伤效果,之后,以杀伤效果较好的肝脏裂解物为探针,以亲和淘洗方法筛选日本血吸虫成虫(44d)噬菌体展示cDNA文库。经三轮筛选后,随机挑取92个噬菌体克隆进行序列测定及生物信息学分析,获得了19个有效EST序列,其中15条EST序列与已知基因或表达序列标签同源,4个EST序列与已知基因或表达序列标签均无同源性,为新的表达序列标签。对19个有效EST编码蛋白的功能预测结果显示,其中6个为卵壳蛋白(Eggshell protein)相关基因,10个EST及其编码蛋白的功能与血吸虫基因表达调控及蛋白质加工修饰相关。研究结果提示,东方田鼠直接或间接地影响血吸虫基因表达,进而影响血吸虫的发育,可能是东方田鼠具有抗血吸虫特性的一个重要分子机理。Microtus fortis （M.fortis） is naturely resisitent to Schistosoma japonicum （S.japonicum）. In order to identify the genes of S japonicu related to the Schistosoma-resistance of M.fortis and expatiate its native resistance mechanism, the Schistosomula-killing effects of tissue extracts of M.fortis were compared and the homogenate supematant of the liver was screened against a phage display cDNA library of S.japonicum. A total of 92 positive clones picked at random were sequenced and 19 ESTs including 4 unreported ESTs were obtained. Sequence analysis indicated that 6 ESTs represented eggshell protein encoding gene and 10 ESTs were involved in the regulation of gene expression and protein modification of Schistosoma japonicum. These results suggested that M.fortis could influence the gene expression of S.japonicum and the body developmentof the parasite, which could be an important mechanism of Schistosoma-resistance of M.fortis.

关 键 词：日本血吸虫 东方田鼠 筛选 噬菌体展示cDNA文库 

分 类 号：S852.735[农业科学—基础兽医学]