secondary chromosomal attachment site and tandem integration of the mobilizable salmonella genomic island 1二次染色体附件网站和串联整合可动员的沙门氏菌基因组岛1.pdfVIP

Secondary Chromosomal Attachment Site and Tandem Integration of the Mobilizable Salmonella Genomic Island 1 ˆ 1 2 2 1 Benoıt Doublet *, George R. Golding , Michael R. Mulvey , Axel Cloeckaert ´ 1 INRA, UR1282, Infectiologie Animale et Sante Publique, Nouzilly, France, 2 National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, Manitoba, Canada Abstract Background: The Salmonella genomic island 1 is an integrative mobilizable element (IME) originally identified in epidemic multidrug-resistant Salmonella enterica serovar Typhimurium (S. Typhimurium) DT104. SGI1 contains a complex integron, which confers various multidrug resistance phenotypes due to its genetic plasticity. Previous studies have shown that SGI1 integrates site-specifically into the S. enterica, Escherichia coli, or Proteus mirabilis chromosome at the 39 end of thdF gene (attB site). Methodology/Principal Findings: Here, we report the transfer of SGI1 to a DthdF mutant of S. Typhimurium LT2. In the absence of thdF, the frequency of transconjugant formation was reduced by around thirty times of magnitude. Through DNA sequencing SGI1 was shown to integrate specifically into a secondary attachment site (2nd attB), which is located in the intergenic region between the chromosomal sodB and purR genes. At this 2nd attB site, we found that a significant fraction of SGI1 transconjugants (43% of wild type and 100% of DthdF mutant) contained tandem SGI1 arrays. Moreover, in wild type S. Typhimurium LT2 transconjugants, SGI1 integrated into both attachment sites, i.e., thdF and sodB-purR. The formation of SGI1 tandem arrays occurred in both specific attB sites. There was heterogeneity in the size of the SGI1 tandem arrays