PMID:4599365

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Citation

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

Abstract

No abstract in PubMed

Links

PubMed

Keywords

Centrifugation, Density Gradient; Chromosome Mapping; DNA Replication; Drug Resistance, Microbial; Escherichia coli; Extrachromosomal Inheritance; Gene Frequency; Mutation; Species Specificity; Streptomycin; Suppression, Genetic; Temperature; Thymine; Tritium

Main Points of the Paper

  • Hfr strains suppress initiation of DNA replication mutants via integrative suppression, giving rise to temperature-resistant revertants
  • The presence of other plasmids affect the reversion rate in Hfr strains
    • Multiple excisions and integrations may take place- the presence of other plasmids may be blocking proper sites for F factor.
  • F-like but not I-like plasmids support integrative suppression in dnaA strains defective for initiation of DNA replication, not dnaC mutants
  • Non-transmissible ColE1 and ColE2 plasmids do not cause integrative suppression
  • Most of the time the entire plasmid is integrated into the chromosome of the suppressed revertants
    • Hemolytic factor integrates only part of the plasmid while the other part remains autonomous
  • Autonomous ColE1 plasmid can only be maintained in limited copies when the chromosome is being replicated under integrative suppression
  • Autonomous R1 factor replication continues under integrative suppression

Materials and Methods Used

  • Integrative Suppression
  • EtBr sensitivity assay
  • Monitor growth rate
  • Thymidine incorporation to measure DNA synthesis rate

Phenotype Annotations

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Species Taxon ID Strain Gene (if known) OMP Phenotype Details Evidence Notes

CRT-46

streptomycin resistant

Resistance to

Plating Assay

CRT-46

abolished lactose utilization

Metabolic Activity

Plating Assay

CRT-46

threonine auxotroph

Growth

Plating Assay

CRT-46

abolished lactose utilization

Metabolic Activity

Plating Assay

CRT-46

leucine auxotroph

Growth

Plating Assay

CRT-46

isoleucine auxotroph

Growth

Plating Assay

CRT-46

thiamine auxotroph

Growth

Plating Assay

CRT-46

thymine auxotroph

Growth

Plating Assay

Escherichia coli

Hfr-120/6

thiamine auxotroph

Growth

Plating Assay

Escherichia coli

Hfr-120/6

thymine auxotroph

Growth

Plating Assay

Escherichia coli

PC2

thymine auxotroph

Growth

Plating Assay

Escherichia coli

PC2

leucine auxotroph

Growth

Plating Assay

Escherichia coli

PC2

streptomyin resistant

Escherichia coli

NCBI:

Hfr-120/6

temperature sensitive defect in initiation of DNA replication


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Notes

References

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