Isolation and characterization of a temperature-sensitive generalized transducing bacteriophage for Vibrio cholerae

CP-T1 is the only described generalized transducing bacteriophage for the i ntestinal pathogen Vibrio cholerae. yet many of its basic biological parame ters remain unknown. Due to low frequencies of transduction and pseudolysog en formation, CP-T1 has not been widely used as a genetic tool. To overcome these limitations, we have isolated a conditional mutant of CP-T1 that exh ibits temperature-sensitive plaque formation. Several biological properties of CP-T1ts were determined, including its restrictive temperature, adsorba nce profile to host cells, burst time, and burst sire. Based on these prope rties, an optimized transduction protocol was designed which resulted in se veral fold higher transduction frequencies for a variety of genetic markers from a number of chromosomal loci. Generalized transduction was also demon strated between classical and El Tor biotype strains of V. cholerae. (C) 20 01 Elsevier Science B.V. All rights reserved.