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

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.