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Multiple copies of MATE elements support autonomous plasmid replication in Aspergillus nidulans
A Aleksenko1, D Gems, J Clutterbuck
1Division of Molecular Genetics, University of Glasgow, UK.
Molecular Microbiology
|April 1, 1996
Summary
The AMA1 sequence, featuring MATE elements, enhances plasmid replication and transformation in Aspergillus nidulans. Multiple copies of repeats improve replication, while single copies cause rearrangements and stable concatenates for higher transformation efficiency.
Area of Science:
- Molecular Biology
- Mycology
- Genetics
Background:
- The AMA1 sequence from Aspergillus nidulans functions as a plasmid replicator and transformation enhancer.
- It consists of two inverted repeats (MATE elements) and a central unique spacer.
Purpose of the Study:
- To investigate the functional elements within AMA1 responsible for plasmid replication and transformation enhancement.
- To determine the minimal sequences and configurations required for these functions.
Main Methods:
- Subclone analysis of the AMA1 sequence.
- Transformation assays in Aspergillus nidulans.
- Analysis of plasmid integration and stability.
Main Results:
- The complete inverted duplication, not the central spacer, is essential for efficient plasmid replication.
- AT-rich regions within the inverted repeats are critical for transformation efficiency.
- Two or more repeat copies, in any orientation, confer replicator function.
- Single MATE elements increase transformation frequency but cause rearrangements and concatenation.
- Multimeric concatenates exhibit enhanced mitotic stability and higher transformation efficiency.
Conclusions:
- The MATE elements within AMA1 are key functional units for replication and transformation.
- Concatenation of AMA1-containing vectors can lead to amplified DNA-like structures with high transformation potential.
- AMA1 offers a system for studying DNA amplification and improving gene delivery in fungi.