牛布鲁菌强毒株544与猪型疫苗株S2基因组差异筛选与鉴定

Screening and identification of the differential genes of genomic DNAs between bovine Brucella virulent strain 544 and vaccine strain S2

应用代表性差异分析技术(Representational difference analysis,RDA)对流产布鲁菌强毒株544和疫苗株S2进行基因组差异分析。经过3轮差减杂交,对差异片段进行Southern-blot验证,BLAST分析和PCR鉴定,最终获得疫苗株S2特有的3条差异片段与已测序的所有猪布鲁菌(Brucella suis)同源性均为100%,只在弱毒株S2基因组中检测出,而在强毒株544中不存在,为建立布鲁菌自然感染菌与疫苗株的基因鉴别诊断方法奠定了基础。

Using representational difference analysis（RDA） to determine the differential genomic DNA between Brucella abortus virulent strain 544 and vaccine strainS2.After three rounds of subtractive hybridization and Southern blot verification,differential fragments were analyzed by BLAST and identified by PCR,3 differential fragments with 100% homology with the sequenced whole Brucella suis,only exist in attenuated strain S2 but were not detected in virulent strain 544 by PCR.It is possible these 3 differential fragments can be used to establish the genic diagnosis to discriminate the Brucella infection from vaccination.