猪肺炎支原体诱导家蝇幼虫抑制性消减文库的构建  

Construction of the suppression subtractive hybridization library induced by Mycoplasma pneumonia

作　　者：王东方[1] 张惠[1] 马红霞[1,2] 高云航[1] 段永杰[1] 王莹[1] 卞璐[1] 王艳芳[3] 

机构地区：[1]吉林农业大学动物科技学院,吉林长春130118 [2]吉林农业大学动物生产与产品质量安全教育部重点实验室,吉林长春130118 [3]吉林农业大学生物反应器与药物开发教育部工程研究中心,吉林长春130118

出　　处：《中国兽医学报》2013年第3期358-361,共4页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(30940055;31140026);吉林省重大科技攻关项目(09ZDGG008);吉林省教育厅"十一五"科学技术研究项目(吉教科200949);吉林省世行贷款农产品质量安全项目(2011-Y05);2010年度教育部新世纪优秀人才计划项目(CET-10-0174)

摘　　要：采用抑制性消减杂交技术(suppression subtractive hybridization,SSH)构建猪肺炎支原体(Mycoplasma hyo-pneumonia,Mhp)诱导家蝇幼虫后产生差异表达基因的消减cDNA文库,并利用反向Northern杂交技术获得差异基因。结果表明,随机挑取的阳性克隆的大小均为200～750bp,通过斑点杂交技术共筛选出186个差异基因片段,对其克隆、测序及同源性分析后鉴定出20种编码蛋白的基因片段和21个无同源序列。家蝇幼虫经猪肺炎支原体诱导后产生与抗病原体相关的基因及大量的未知基因。In order to get stronger antimicrobial peptides to replace antibiotic drugs, this experi- ment constructed subtractive cDNA libraries about Musca domestica larvae induced by Mycoplas- rna hyopneumonia by suppression subtractive hybridization, and got the differential gene by re- verse northern blot. The results showed that, the randomly selected positive clones were all in 200-750 bp, 186 differential genes fragments screened by dot blot were cloned,sequenced and ho- mology analysis,identified 20 protein genes fragment and 21 without homologous sequence. Musca domestica larvae induced by MHP produced anti-pathogen genes and a large number of unknown genes.

关 键 词：猪肺炎支原体 家蝇 抑制性消减杂交 反向Northern 

分 类 号：S852.62[农业科学—基础兽医学]