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[RP4 factor integration with E. coli chromosome].

V N Danilevich, Iu G Stepanshin, N V Volozhantsev

    Antibiotiki
    |April 1, 1978
    PubMed
    Summary
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    High-frequency integration of R-factor RP4 into E. coli chromosomes occurred when the R-factor contained transposon Tn1 and the chromosome already had Tn1. This suggests a recA-dependent recombination mechanism involving Tn1.

    Area of Science:

    • Microbiology
    • Molecular Biology
    • Bacterial Genetics

    Background:

    • R-factors, like RP4, are plasmids that can integrate into bacterial chromosomes.
    • Transposons, such as Tn1, are mobile genetic elements that can facilitate integration.
    • The recA gene is crucial for homologous recombination in E. coli.

    Purpose of the Study:

    • To investigate the mechanism of R-factor RP4 integration into the E. coli chromosome.
    • To determine the role of transposon Tn1 and the recA gene in this integration process.

    Main Methods:

    • Utilized a replication thermosensitive mutant of R-factor RP4 (pEG1).
    • Studied integration frequencies in E. coli strains JC411 (with pre-inserted Tn1) and JC1553 (recA defective).
    • Compared integration rates in the presence and absence of Tn1 on the bacterial chromosome.

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    Main Results:

    • Integration frequency of pEG1 was significantly higher in JC411 (carrying Tn1) compared to E. coli without Tn1.
    • Integration frequency of pEG1 into recA-defective JC1553 was very low (<2.10^-5) and independent of Tn1 presence.
    • These findings suggest a recA-dependent mechanism for RP4 integration.

    Conclusions:

    • R-factor RP4 integration into the E. coli chromosome likely occurs via recA-dependent recombination.
    • The presence of transposon Tn1 on both the R-factor and the chromosome greatly enhances integration frequency.
    • This mechanism involves homologous recombination between Tn1 elements.