Frequency of DNA transfer of phage T4 as determined by CPRINS-FISH

Range 9e-4 - 3e-3 gene/PFU
Organism Bacteria Escherichia coli
Reference Kenzaka T, Tani K, Sakotani A, Yamaguchi N, Nasu M. High-frequency phage-mediated gene transfer among Escherichia coli cells, determined at the single-cell level. Appl Environ Microbiol. 2007 May73(10):3291-9.PubMed ID17384307
Method Purification of bacteriophages. Sequencing of isolated phage EC10. Plaque assays. Transduction and DNA transfer experiments. Viabilities of recipient cells determined by Direct Viable Counting (DVC). (cycling primed in situ amplification-fluorescent in situ hybridization [CPRINS-FISH]). Epifluorescence microscopy. Genetic characterization of transductants. PCR amplification of the unknown region next to the (beta lactamase gene-bla) sequence on transductants and sequencing. Examination of integration of bla gene into the phage genome.
Comments PFU=Plaque Forming Unit. Transduction frequencies determined by conventional plating were 1x10^(-8) - 4x10^(-8) gene/PFU
Entered by Uri M
ID 104446