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The mini-F primary origin. Sequence analysis and multiple activities.

D Lane, D Hill, P Caughey

    Journal of Molecular Biology
    |December 5, 1984
    PubMed
    Summary
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    The mini-F origin of replication (ori-1) sequence was determined, revealing a protein essential for its activity. Replication requires specific DNA sequences and a trans-acting gene product, likely the E protein.

    Area of Science:

    • Molecular Biology
    • Genetics
    • Microbiology

    Background:

    • The F genetic map details crucial regions for mini-F plasmid replication.
    • Understanding replication origins is key to plasmid biology and genetic engineering.

    Purpose of the Study:

    • To determine the sequence of the mini-F primary origin of replication (ori-1).
    • To identify genetic elements and factors necessary for ori-1 activity.

    Main Methods:

    • DNA sequencing of an 897 base-pair fragment containing ori-1.
    • Functional analysis of ori-1 in DNA polymerase I-dependent replicons.

    Main Results:

    • Identified an open reading frame likely coding for a replication-associated protein (C protein).

    Related Experiment Videos

  • ori-1 replication necessitates adjacent mini-F sequences (43.0-43.9 kb) in cis.
  • Replication also requires a trans-acting gene product, likely the E protein.
  • Mapped a pif gene expression control sequence and a recombination site near the C (pifC) gene.

    • Conclusions:

    • The sequenced fragment contains essential elements for mini-F replication.
    • Replication is a complex process involving cis- and trans-acting factors.
    • Identified regulatory elements for gene expression and recombination within the mini-F replication region.