Journal
JOURNAL OF MICROBIOLOGICAL METHODS
Volume 46, Issue 3, Pages 217-225Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/S0167-7012(01)00276-7
Keywords
Vibrio bacteriophage; generalized transduction; Vibrio cholerae; CP-T1ts
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Funding
- NIAID NIH HHS [AI40262] Funding Source: Medline
- NIDDK NIH HHS [P30 DK34928] Funding Source: Medline
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CP-T1 is the only described generalized transducing bacteriophage for the intestinal pathogen Vibrio cholerae. yet many of its basic biological parameters remain unknown. Due to low frequencies of transduction and pseudolysogen 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 exhibits temperature-sensitive plaque formation. Several biological properties of CP-T1ts were determined, including its restrictive temperature, adsorbance profile to host cells, burst time, and burst sire. Based on these properties, an optimized transduction protocol was designed which resulted in several fold higher transduction frequencies for a variety of genetic markers from a number of chromosomal loci. Generalized transduction was also demonstrated between classical and El Tor biotype strains of V. cholerae. (C) 2001 Elsevier Science B.V. All rights reserved.
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