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Related Experiment Video

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CAPRRESI: Chimera Assembly by Plasmid Recovery and Restriction Enzyme Site Insertion

CAPRRESI: Chimera Assembly by Plasmid Recovery and Restriction Enzyme Site Insertion

Published on: June 25, 2017

A Type III restriction-modification system in Mycoplasma mycoides subsp. capri.

Mikkel A Algire¹, Michael G Montague, Sanjay Vashee

¹The J. Craig Venter Institute, 9704 Medical Center Drive, Rockville, MD 20850, USA. malgire@jcvi.org

|November 17, 2012

Summary

Mycoplasma mycoides subsp. capri possesses a Type III restriction-modification system (MmyCI). Its methyltransferase gene expression is phase-variable due to AG repeats, affecting MmyCI activity.

Keywords:

Type III methyltransferase mycoplasma phase variation

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Area of Science:

Microbiology
Genomics
Molecular Biology

Background:

The genome sequencing of Mycoplasma mycoides subsp. capri identified a Type III restriction-modification system, MmyCI.
The methyltransferase (M.MmyCI) component of MmyCI recognizes and methylates the 5'-TGAG-3' sequence.

Purpose of the Study:

To investigate the MmyCI system in Mycoplasma mycoides subsp. capri.
To determine the mechanism behind the observed lack of MmyCI activity in certain strains.

Main Methods:

Genome sequencing and analysis.
Gene expression analysis focusing on the methyltransferase gene.
Investigation of DNA repeats and their effect on protein translation.

Main Results:

The methyltransferase gene contains a variable number of AG dinucleotide repeats.
A strain with 12 AG repeats exhibited translational termination, resulting in no MmyCI activity.
A clone with 10 AG repeats allowed for in-frame gene expression and MmyCI activity.

Conclusions:

The number of AG repeats in the MmyCI methyltransferase gene influences its expression.
Phase variation in AG repeat number likely controls MmyCI activity in Mycoplasma mycoides subsp. capri.