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Journal of Bacteriology, June 2000, p. 3559-3571, Vol. 182, No. 12
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Characterization of a Bacteroides Mobilizable
Transposon, NBU2, Which Carries a Functional Lincomycin
Resistance Gene
Jun
Wang,
Nadja B.
Shoemaker,
Gui-Rong
Wang, and
Abigail A.
Salyers*
Department of Microbiology, University of
Illinois, Urbana, Illinois 61801
Received 20 October 1999/Accepted 24 March 2000
The mobilizable Bacteroides element NBU2 (11 kbp) was
found originally in two Bacteroides clinical isolates,
Bacteroides fragilis ERL and B. thetaiotaomicron DOT. At first, NBU2 appeared to be very similar
to another mobilizable Bacteroides element, NBU1, in a
2.5-kbp internal region, but further examination of the full DNA
sequence of NBU2 now reveals that the region of near identity between
NBU1 and NBU2 is limited to this small region and that, outside this
region, there is little sequence similarity between the two elements.
The integrase gene of NBU2, intN2, was located at one end
of the element. This gene was necessary and sufficient for the
integration of NBU2. The integrase of NBU2 has the conserved amino
acids (R-H-R-Y) in the C-terminal end that are found in members of the
lambda family of site-specific integrases. This was also the only
region in which the NBU1 and NBU2 integrases shared any similarity
(28% amino acid sequence identity and 49% sequence similarity).
Integration of NBU2 was site specific in Bacteroides
species. Integration occurred in two primary sites in B. thetaiotaomicron. Both of these sites were located in the 3' end
of a serine-tRNA gene NBU2 also integrated in Escherichia coli, but integration was much less site specific than in
B. thetaiotaomicron. Analysis of the sequence of NBU2
revealed two potential antibiotic resistance genes. The amino acid
sequences of the putative proteins encoded by these genes had
similarity to resistances found in gram-positive bacteria. Only one of
these genes was expressed in B. thetaiotaomicron, the
homolog of linA, a lincomycin resistance gene from
Staphylococcus aureus. To determine how widespread elements related to NBU1 and NBU2 are in Bacteroides species, we
screened 291 Bacteroides strains. Elements with some
sequence similarity to NBU2 and NBU1 were widespread in
Bacteroides strains, and the presence of
linAN in Bacteroides strains was
highly correlated with the presence of NBU2, suggesting that NBU2 has
been responsible for the spread of this gene among
Bacteroides strains. Our results suggest that the
NBU-related elements form a large and heterogeneous family, whose
members have similar integration mechanisms but have different target
sites and differ in whether they carry resistance genes.
*
Corresponding author. Mailing address: Department of
Microbiology, B103 CLSL, 601 S. Goodwin, Urbana, IL 61801. Phone: (217) 333-7378. Fax: (217) 244-8485. E-mail: abigails{at}uiuc.edu.
Journal of Bacteriology, June 2000, p. 3559-3571, Vol. 182, No. 12
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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