3种保虫宿主日本血吸虫特征性差异表达基因的筛选与验证  

Screening and verification on characteristic differentially expressed genes of Schistosoma japonicum from three reservoir hosts

作　　者：杨健美[1] 石耀军[1] 冯新港[1] 傅志强[1] 苑纯秀[1] 刘金明[1] 洪炀[1] 李浩[1] 陆珂[1] 林矫矫[1] 

机构地区：[1]中国农业科学院上海兽医研究所、农业部动物寄生虫学重点实验室,上海200241

出　　处：《中国血吸虫病防治杂志》2012年第3期279-283,共5页Chinese Journal of Schistosomiasis Control

基　　金：国家重大科技专项(2012ZX10004220-008,2008ZX10004-011);公益性行业(农业)科研专项(200903036)

摘　　要：目的筛选日本血吸虫3种重要保虫宿主来源虫体的特征性差异表达基因,寻找可以用于鉴别这3种宿主来源虫体的遗传标记。方法日本血吸虫尾蚴人工感染黄牛、水牛和山羊,收集感染后49 d成虫,分别抽提RNA,逆转录成cD NA,体外转录为cRNA并进行片段化;采用定制的血吸虫芯片分别进行杂交,筛选每种宿主来源虫体的特征性差异表达基因。采用实时定量PCR法对所筛选差异表达基因的表达水平做进一步测定,验证芯片数据的结果,并对这些特征性差异表达基因进行验证。结果通过芯片杂交,分析筛选获得实验黄牛组来源虫体特征性差异表达基因3个,水牛组来源虫体4个,山羊组来源虫体7个。其中黄牛组2个、水牛组3个、山羊组5个特征性差异表达基因在另一批次3种宿主来源虫体的重复试验中得到验证。结论芯片杂交筛选出10个3种宿主来源虫体的特征性差异表达基因,这些基因可能可以作为这3种不同宿主来源虫体的遗传标记。Objective To get the characteristic differentially expressed genes of Schistosomajaponicum from three important reservoir hosts ： yellow cattle, water buffalo and goat, so as to find the genetic markers to identify the various sources of the para- site reservoir hosts. Methods The 49 d worms were collected from artificially infected animals, and the total RNA （s） of worms were extracted and reverse-transcripted to cDNA, and then hybridized with custom-built microarray to screen characteristic differ- entially expressed genes of every host, and the microarray results were validated by the real-time PCR method. Results From re- sults of microarray, we got 3 characteristic differentially expressed genes of S. japonicum from yellow cattle, 4 from water buffalo and 7 from goat. We verified schistosome samples from three reservoir hosts in another experiment, the results showed that 2 in yel- low cattle, 3 in water buffalo, and 5 in goat were verified to be consistent with microarray results. Conclusions The ten charac- teristic differentially expressed genes of S. japonicum from three reservoir hosts screened by microarray might be used as genetic markers to identify the various sources of reservoir hosts for S. japonicum.

关 键 词：日本血吸虫 保虫宿主 芯片 特征性差异表达基因 

分 类 号：R383.24[医药卫生—医学寄生虫学]