![]() Microbes can acquire new genetic material from neighboring organisms via a process termed horizontal gene transfer (HGT). Thus, beyond the genetic acquisition of novel DNA sequences, NT can also promote the nongenetic inheritance of traits during this conserved mechanism of horizontal gene transfer. We show that the combination of these properties results in the nongenetic transfer of gene products within transformed populations, which can support phenotypic inheritance of antibiotic resistance in both V. Our results directly demonstrate (1) that transforming DNA efficiently integrates into the genome as single-stranded DNA, (2) that the resulting heteroduplexes are resolved by chromosome replication and segregation, and (3) that integrated DNA is rapidly expressed prior to cell division. Here, we develop a cell biological approach to characterize the spatial and temporal dynamics of homologous recombination during NT in Vibrio cholerae. Natural transformation (NT) is a major mechanism of horizontal gene transfer in microbial species that promotes the spread of antibiotic resistance determinants and virulence factors. Data are from three independent experiments and n = 104 and n = 101 GFP positive cells analyzed, for B and C respectively. Cells in C also contained a Δ mutS mutation to inactivate the mismatch repair system. Histogram of the time to tDNA-derived GFP expression relative to chromosome replication (which occurs at t = 0) for repair of ( B) an 87bp deletion in gfp or ( C) two point mutations that introduced a premature stop codon into gfp. Experiments were imaged every 3 mins for 5 hours. ![]() Cells were then transformed with tDNA to delete the parS1 site and repair the mutation in the gfp gene. The parS1 site sits upstream of an inactivated gfp gene (via an 87bp deletion of the 5’ end 1 of the gfp gene or two point mutations that introduce a premature stop codon into gfp as indicated below). Cells contained parS1 (red ParB1) and parS2 (cyan ParB2) sites in close proximity on the chromosome. ![]() 5: Figure S5 – Expression of newly integrated DNA occurs rapidly following chromosome replication, Related to Figure 4.
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