Category:Integrative Suppression

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Purpose

  • Suppression of mutants defective in initiation of DNA replication due to a mutation in dnaA[1][2]
  • Isolation of mutants defective in initiation of DNA replication[3]

Technical Details

  • To suppress dnaA-based initiation of DNA replication mutants

At restrictive temperature, the DnaA mutant protein does not allow for initiation of DNA replication at oriC. This defect is suppressed by initiation from the integrated plasmid's origin through which replication of the chromosome proceeds. This mechanism results in temperature-resistant, DnaA-independent initiation of DNA replication.[1][2]

Materials Needed

  • Various replicons can be used
    • F and F' factors[1]
    • R factors[4]
    • bacteriophage P1 and P7[5]
    • bacteriophage P2 sig5 (mutant bacteriophage of P2)[6][7]

Methods

Examples

Notes

References

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  1. ↑ 1.0 1.1 1.2 Nishimura, Y et al. (1971) Chromosome replication in Escherichia coli. IV. Control of chromosome replication and cell division by an integrated episome. J. Mol. Biol. 55 441-56 PubMed OMPwiki page
  2. ↑ 2.0 2.1 Goebel, W (1974) Integrative suppression of temperature-sensitive mutants iwth a lesion in the initiation of DNA replication. Replication of autonomous plasmids in the suppressed state. Eur. J. Biochem. 43 125-30 PubMed OMPwiki page
  3. ↑ Schaus, N et al. (1981) Isolation and characterization of amber mutations in gene dnaA of escherichia coli K-12. J. Bacteriol. 145 904-13 PubMed OMPwiki page
  4. ↑ Nishimura, A et al. (1973) Isolation of Hfr strains from R+ and ColV2+ strains of Escherichia coli and derivation of an R'lac factor by transduction. J. Bacteriol. 116 1107-12 PubMed OMPwiki page
  5. ↑ Chesney, RH & Scott, JR (1978) Suppression of a thermosensitive dnaA mutation of Escherichia coli by bacteriophage P1 and P7. Plasmid 1 145-63 PubMed OMPwiki page
  6. ↑ Kuempel, PL et al. (1978) Chromosome replication in an Escherichia coli dnaA mutant integratively suppressed by prophage P2. J. Bacteriol. 134 902-12 PubMed OMPwiki page
  7. ↑ Lindahl, G et al. (1971) On the process of cellular division in Escherichia coli: replication of the bacterial chromosome under control of prophage P2. Proc. Natl. Acad. Sci. U.S.A. 68 2407-11 PubMed OMPwiki page

Pages in category "Integrative Suppression"

The following 2 pages are in this category, out of 2 total.

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