牛多杀性巴氏杆菌诱导家蝇幼虫抑制性消减文库的构建  

Construction of SSH library of Musca domestica larvae upon infection with Pasteurella mutocida of bovine

作　　者：王莹[1] 高云航[2] 马红霞[2] 裴志花[2] 唐艳[2] 

机构地区：[1]吉林农业大学生命科学学院,吉林长春130118 [2]吉林农业大学动物科技学院/动物生产与产品质量安全教育部重点实验室,吉林长春130118

出　　处：《中国兽医学报》2013年第5期697-700,共4页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31140026;31272611);吉林省科技发展计划资助项目(20111820);教育部新世纪优秀人才资助项目(NCET-10-0174);吉林省科技发展计划资助项目(20120229);吉林省世行贷款农产品质量安全资助项目(2011-Y05)

摘　　要：采用牛多杀性巴氏杆菌针刺法诱导3日龄家蝇幼虫,提取诱导后的家蝇幼虫总RNA并分离纯化得到高质量的mRNA。运用抑制性消减杂交(Suppression subtractive hybridization,SSH)技术构建家蝇幼虫差异表达的cD-NA消减文库,并利用反向Northern杂交技术筛选差异表达基因。结果表明,随机挑取的阳性克隆片段大小均在200～750bp,通过反向Northern杂交共筛选出183个差异表达基因,经BLAST比对分析后鉴定出大量与家蝇免疫防御功能相关的基因和一部分无同源性基因。Three-day-old Musca dornestica larvae were pricked with a needle dipped in Pasteurella rnultocida of Bovine. The total RNA was extracted from Musca domestica larvae,and we got the high quality mRNA after isolation and purification. Suppression subtractive hybridization（SSH） was performed to generate a subtracted cDNA library,and the differentially expressed genes were screened by reverse Northern hybridization technology. The results showed that the sizes of positive clonal fragments that picked randomly were between 200bp and 750bp and 183 genes differentially expressed were screened by reverse Northern hybridization technology. A large number of genes that associated with the immune defensive of Musca domestica and a portion of the non-homologous genes were identified by BLAST.

关 键 词：家蝇幼虫 牛多杀性巴氏杆菌 抑制性消减杂交 反向Northern BLOT 

分 类 号：S852.61[农业科学—基础兽医学]