Tn1546 is part of a larger plasmid-encoded genetic unit horizontally disseminated among clonal Enterococcus faecium lineages.
OBJECTIVE
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).


METHODS
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.


RESULTS
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.


CONCLUSIONS
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.
Sletvold
H
H
Department of Pharmacy, Faculty of Medicine, University of Tromsø, 9037 Tromsø, Norway.
Johnsen
P J
PJ
Wikmark
O-G
OG
Simonsen
G S
GS
Sundsfjord
A
A
Nielsen
K M
KM
D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics*; D019729:Carbon-Oxygen Ligases / Q000235:genetics*; D016386:Chromosome Walking; D016000:Cluster Analysis; D017931:DNA Primers / Q000235:genetics; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D016984:Enterococcus faecium / Q000235:genetics* / Q000302:isolation & purification; D005602:France; D023061:Gene Order; D022761:Gene Transfer, Horizontal; D016908:Gram-Positive Bacterial Infections / Q000382:microbiology / Q000662:veterinary; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010957:Plasmids; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017385:Sequence Homology
D001426:Bacterial Proteins; D017931:DNA Primers; D004251:DNA Transposable Elements; D004269:DNA, Bacterial; C071437:VanA ligase, Bacteria; D019729:Carbon-Oxygen Ligases