Phage-mediated horizontal gene transfer of both prophage and heterologous DNA by ϕBB-1, a bacteriophage of Borrelia burgdorferi.
Horizontal gene transfer (HGT) in Borrelia burgdorferi, the Lyme disease agent, is likely mediated by bacteriophage. Studies of the B. burgdorferi phage, ϕBB-1 and its role in HGT have been hindered by the lack of an assay for readily characterizing phage-mediated DNA movement (transduction). Here we describe an in vitro assay in which a clone of B. burgdorferi strain CA-11.2A encoding kanamycin resistance on a ϕBB-1 prophage is co-cultured with different clones encoding gentamicin resistance on a shuttle vector; transduction is monitored by enumerating colonies selected in the presence of both kanamycin and gentamicin. When both clones used in the assay were derived from CA-11.2A, the frequency of transduction was 1.23 × 10-6transductants per cell, and could be increased 5-fold by exposing the phage-producing strain to 5% ethanol. Transduction was also demonstrated between the CA-11.2A clone and clones of both high-passage B. burgdorferi strain B31 and low-passage, virulent B. burgdorferi strain 297, although with lower transduction frequencies. The transductant in the 297 background produced phage capable of transducing another B. burgdorferi clone: this is the first experimental demonstration of transduction from a clone of a virulent strain. In addition to prophage DNA, small Escherichia coli-derived shuttle vectors were also transduced between co-cultured B. burgdorferi strains, suggesting both a broad role for the phage in the HGT of heterologous DNA and a potential use of the phage as a molecular tool. These results enhance our understanding of phage-mediated transduction as a mechanism of HGT in the Lyme disease spirochetes. Furthermore, the reagents and techniques developed herein will facilitate future studies of phage-mediated HGT, especially within the tick vector and vertebrate host.
Department of Biomedical Sciences, Quinnipiac University, Hamden, CT 06518, USA christian.eggers@quinnipiac.edu.
Christian H
CH
Eggers
Department of Biomedical Sciences, Quinnipiac University, Hamden, CT 06518, USA.
Carlie M
CM
Gray
Department of Biomedical Sciences, Quinnipiac University, Hamden, CT 06518, USA.
Alexander M
AM
Preisig
Department of Biomedical Sciences, Quinnipiac University, Hamden, CT 06518, USA.
Danielle M
DM
Glenn
Department of Biomedical Sciences, Quinnipiac University, Hamden, CT 06518, USA.
Jessica
J
Pereira
Department of Biomedical Sciences, Quinnipiac University, Hamden, CT 06518, USA.
Ryan W
RW
Ayers
Department of Biomedical Sciences, Quinnipiac University, Hamden, CT 06518, USA.
Mohammad
M
Alshahrani
Department of Biomedical Sciences, Quinnipiac University, Hamden, CT 06518, USA.
Christopher
C
Acabbo
Department of Biomedical Sciences, Quinnipiac University, Hamden, CT 06518, USA.
Maria R
MR
Becker
Department of Biomedical Sciences, Quinnipiac University, Hamden, CT 06518, USA.
Kimberly N
KN
Bruenn
Department of Biomedical Sciences, Quinnipiac University, Hamden, CT 06518, USA.
Timothy
T
Cheung
Department of Biomedical Sciences, Quinnipiac University, Hamden, CT 06518, USA.
Taylor M
TM
Jendras
Department of Biomedical Sciences, Quinnipiac University, Hamden, CT 06518, USA.
Aron B
AB
Shepley
Department of Biomedical Sciences, Quinnipiac University, Hamden, CT 06518, USA.
John T
JT
Moeller
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001435:Bacteriophages / Q000187:drug effects / Q000502:physiology*; D025065:Borrelia burgdorferi / Q000187:drug effects / Q000235:genetics* / Q000821:virology*; D004247:DNA; D004269:DNA, Bacterial; D000431:Ethanol / Q000494:pharmacology; D022761:Gene Transfer, Horizontal; D006801:Humans; D008193:Lyme Disease / Q000382:microbiology; D008826:Microbial Sensitivity Tests
D000900:Anti-Bacterial Agents; D004269:DNA, Bacterial; D000431:Ethanol; D004247:DNA
Borrelia burgdorferi; Lyme disease; bacteriophage; horizontal gene transfer; transduction