Affiliation:
1. Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamada-oka, Osaka 565-0871, Japan
2. Faculty of Pharmacy, Osaka Ohtani University, 3-11-1 Nishikiori-kita, Tondabayashi 584-8540, Japan
Abstract
ABSTRACT
Recent whole-genome analysis suggests that lateral gene transfer by bacteriophages has contributed significantly to the genetic diversity of bacteria. To accurately determine the frequency of phage-mediated gene transfer, we employed cycling primed in situ amplification-fluorescent in situ hybridization (CPRINS-FISH) and investigated the movement of the ampicillin resistance gene among
Escherichia coli
cells mediated by phage at the single-cell level. Phages P1 and T4 and the newly isolated
E. coli
phage EC10 were used as vectors. The transduction frequencies determined by conventional plating were 3 × 10
−8
to 2 × 10
−6
, 1 × 10
−8
to 4 × 10
−8
, and <4 × 10
−9
to 4 × 10
−8
per PFU for phages P1, T4, and EC10, respectively. The frequencies of DNA transfer determined by CPRINS-FISH were 7 × 10
−4
to 1 × 10
−3
, 9 × 10
−4
to 3 × 10
−3
, and 5 × 10
−4
to 4 × 10
−3
for phages P1, T4, and EC10, respectively. Direct viable counting combined with CPRINS-FISH revealed that more than 20% of the cells carrying the transferred gene retained their viabilities. These results revealed that the difference in the number of viable cells carrying the transferred gene and the number of cells capable of growth on the selective medium was 3 to 4 orders of magnitude, indicating that phage-mediated exchange of DNA sequences among bacteria occurs with unexpectedly high frequency.
Publisher
American Society for Microbiology
Subject
Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology
Cited by
31 articles.
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