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Determination of the Optimal Chromosomal Location(s) for a DNA Element in Escherichia coli Using a Novel Transposon-mediated Approach
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Host specificity of mollicutes oriC plasmids: functional analysis of replication origin

Carole Lartigue1, Alain Blanchard, Joël Renaudin

  • 1UMR GDPP, INRA-Université Victor Segalen Bordeaux 2, BP 81, 33883 Villenave d'Ornon Cedex, France.

Nucleic Acids Research
|November 7, 2003
PubMed

Insights

Artificial oriC plasmids reveal replication specificity among mollicutes. The dnaA gene region functions as a replication origin, but host factors influence heterologous oriC plasmid replication, indicating complex specificity determinants.

Area of Science:

  • Molecular Biology
  • Microbiology
  • Genetics

Background:

  • Artificial oriC plasmids containing the dnaA gene and DnaA boxes were previously developed for Spiroplasma citri and Mycoplasma pulmonis.
  • Mollicutes, a group of bacteria, possess unique genetic features and are known pathogens.

Purpose of the Study:

  • To investigate the host specificity of oriC plasmids across different mollicute species.
  • To determine if the dnaA gene region serves as a conserved replication origin in mollicutes.
  • To identify factors beyond cis-elements that contribute to oriC plasmid replication specificity.

Main Methods:

  • Construction of artificial oriC plasmids for Mycoplasma mycoides subsp. mycoides LC (MmmLC), M. mycoides subsp. mycoides SC (MmmSC), and Mycoplasma capricolum subsp. capricolum.
  • Transformation experiments using homologous and heterologous oriC plasmids in MmmLC, MmmSC, M. capricolum, S. citri, and M. pulmonis.

Main Results:

  • All five mollicute species were successfully transformed with homologous oriC plasmids, supporting the dnaA gene region as a functional replication origin.
  • Replication of heterologous oriC plasmids varied significantly between species.
  • M. capricolum oriC plasmids failed to replicate in MmmSC and MmmLC, while MmmSC, MmmLC, and S. citri oriC plasmids replicated in M. capricolum.

Conclusions:

  • The dnaA gene region is a functional replication origin in mollicute chromosomes.
  • Host-specific factors, in addition to oriC cis-elements, play a crucial role in determining the replication ability of heterologous oriC plasmids.
  • Mollicute oriC plasmid replication exhibits complex host specificity not solely determined by DNA sequences within the origin.

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