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. 1992 Jan;58(1):187–193. doi: 10.1128/aem.58.1.187-193.1992

High-Frequency Plasmid Transduction by Lactobacillus gasseri Bacteriophage φadh †

R R Raya 1,‡, T R Klaenhammer 1,*
PMCID: PMC195190  PMID: 16348621

Abstract

The temperate bacteriophage φadh mediates plasmid DNA transduction in Lactobacillus gasseri ADH at frequencies in the range of 10-8 to 10-10 transductants per PFU. BglII-generated DNA fragments from phage φadh were cloned into the BclI site of the transducible plasmid vector pGK12 (4.4 kb). Phage φadh lysates induced from Lactobacillus lysogens harboring pGK12 or the recombinant plasmids were used to transduce strain ADH to chloramphenicol resistance. The transduction frequencies of recombinant plasmids were 102- to 105-fold higher than that of native pGK12. The increase in frequency generally correlated with the extent of DNA-DNA homology between plasmid and phage DNAs. The highest transduction frequency was obtained with plasmid pTRK170 (6.6 kb), a pGK12 derivative containing the 1.4- and 0.8-kb BglII DNA fragments of φadh. DNA hybridization analysis of pTRK170-transducing phage particles revealed that pTRK170 had integrated into the φadh genome, suggesting that recombination between homologous sequences present in phage and plasmid DNAs was responsible for the formation of high-frequency transducing phage particles. Plasmid DNA analysis of 13 transductants containing pTRK170 showed that each had acquired intact plasmids, indicating that in the process of transduction a further recombination step was involved in the resolution of plasmid DNA monomers from the recombinant pTRK170::φadh molecule. In addition to strain ADH, pTRK170 could be transduced via φadh to eight different L. gasseri strains, including the neotype strain, F. Gasser 63 AM (ATCC 33323).

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Selected References

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