Department of Pharmacy, Faculty of Medicine, University of Tromsø, 9037 Tromsø, Norway.
J Antimicrob Chemother. 2010 Sep;65(9):1894-906. doi: 10.1093/jac/dkq219. Epub 2010 Jun 17.
To determine the genetic composition of the first VanA-type plasmid (pIP816) reported, which was isolated from a clinical Enterococcus faecium (BM4147) strain in France in 1986, and to reveal the genetic units responsible for the dissemination of the vanA gene cluster by comparisons with current, published and additionally generated vanA-spanning plasmid sequences obtained from a heterogeneous E. faecium strain collection (n = 28).
Plasmid sequences were produced by shotgun sequencing using ABI dye chemistry and primer walking, and were subsequently annotated. Comparative sequence analysis of the vanA region was done with published plasmids, with a partial vanA plasmid (pVEF4) reported here and to >140 kb of sequence obtained from a collection of vanA-harbouring plasmid fragments.
Bioinformatic analyses revealed that pIP816 from 1986 and contemporary vanA plasmids shared a conserved genetic fragment of 25 kb, spanning the 10.85 kb vanA cluster encoded by Tn1546, and that the larger unit is present in both clinical and animal complexes of E. faecium. A new group II intron in pVEF4 was characterized.
Comparative DNA analyses suggest that Tn1546 disseminates in and between clonal complexes of E. faecium as part of a larger genetic unit, possibly as a composite transposon flanked by IS1216 elements.
确定 1986 年从法国临床粪肠球菌(BM4147)菌株中分离出的首个 VanA 型质粒(pIP816)的遗传组成,并通过与当前已发表的和另外生成的来自异质粪肠球菌菌株集(n = 28)的 VanA 基因簇传播相关的遗传单元进行比较,揭示 VanA 基因簇传播的遗传单元。
使用 ABI 染料化学和引物步行进行鸟枪法测序以产生质粒序列,随后进行注释。使用已发表的质粒对 VanA 区进行比较序列分析,本文还报道了部分 VanA 质粒(pVEF4),并对来自 VanA 携带质粒片段集的>140kb 序列进行了分析。
生物信息学分析显示,1986 年的 pIP816 和当代的 VanA 质粒共享 25kb 的保守遗传片段,跨越了由 Tn1546 编码的 10.85kb VanA 簇,而较大的单元存在于粪肠球菌的临床和动物复合体中。pVEF4 中的一个新的组 II 内含子被表征。
比较 DNA 分析表明,Tn1546 作为更大遗传单元的一部分,在粪肠球菌的克隆复合体中传播,可能作为由 IS1216 元件侧翼的复合转座子。