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Related Experiment Videos

Autonomous plasmid replication in Aspergillus nidulans: AMA1 and MATE elements

A Aleksenko1, A J Clutterbuck

  • 1Institute of Genetics and Selection of Industrial Microorganisms, Moscow, Russia.

Fungal Genetics and Biology : FG & B
|June 1, 1997
PubMed
Summary

Filamentous fungi harbor DNA fragments enabling extrachromosomal plasmid replication. Duplication of inefficient replicators enhances their efficiency, suggesting a model for effective plasmid formation via sequence amplification.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Mycology

Background:

  • Extrachromosomal plasmid maintenance in eukaryotes is challenging.
  • Specific DNA fragments from filamentous fungi enable stable plasmid replication.
  • Two classes of sequences in Aspergillus nidulans facilitate plasmid maintenance: replicators and enhancers.

Purpose of the Study:

  • To investigate DNA fragments enabling extrachromosomal plasmid maintenance in fungi.
  • To elucidate the mechanisms behind effective plasmid replication and vector stability.
  • To explore the role of sequence amplification and topoisomerases in plasmid dynamics.

Main Methods:

  • Isolation and characterization of DNA fragments from Aspergillus nidulans.
  • Functional analysis of plasmid replication and maintenance in vivo.

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  • Comparative analysis of DNA amplification mechanisms across eukaryotes.

    • Main Results:

    • Identified effective plasmid replicators (e.g., AMA1) and transformation enhancers (e.g., ANS1, MATEs) in A. nidulans.
    • Observed that enhancer activity leads to vector rearrangement, multimerization, and chromosomal integration.
    • Demonstrated that duplication of inefficient replicators significantly enhances their efficiency in A. nidulans and Schizosaccharomyces pombe.

    Conclusions:

    • A model is proposed where sequence amplification generates effective replicating plasmids.
    • Topoisomerase activity is implicated in the amplification and replication of circular DNA molecules.
    • Replicative vectors hold practical applications for gene cloning and expression studies.