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Updated: May 28, 2025

Site-specific Bacterial Chromosome Engineering: ΦC31 Integrase Mediated Cassette Exchange (IMCE)

Site-specific Bacterial Chromosome Engineering: ΦC31 Integrase Mediated Cassette Exchange (IMCE)

Published on: March 16, 2012

SGI1 excludes IncA and IncC plasmids.

Stephanie J Ambrose¹, Ruth M Hall¹

¹School of Life and Environmental Sciences, The University of Sydney, NSW 2006, Australia.

|February 13, 2025

Summary

This summary is machine-generated.

Integrative mobilizable elements like SGI1 can prevent related plasmids from entering cells. Deleting specific genes in SGI1 variants abolished this plasmid exclusion effect, but the exact mechanism remains unclear.

Keywords:

IncA plasmid IncC plasmid Integrative mobilizable element Plasmid exclusion Plasmid transfer Salmonella genomic island 1

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Area of Science:

Microbiology
Molecular Biology
Genetics

Background:

SGI1 and its variants are integrative mobilizable elements dependent on IncA/IncC plasmids for transfer.
Long-term coexistence of SGI1 and plasmids is unstable, suggesting mechanisms like plasmid exclusion.
Understanding SGI1's effect on plasmid entry is crucial for bacterial genetics and mobile element research.

Purpose of the Study:

To investigate the impact of SGI1 type integrative elements on the initial entry of IncA and IncC plasmids.
To identify the genetic determinants within SGI1 responsible for plasmid exclusion.
To elucidate the mechanism underlying SGI1-mediated plasmid exclusion.

Main Methods:

Utilized two transfer-proficient IncC plasmids and the IncA plasmid RA1.
Constructed and analyzed SGI1 variants (SGI1-I, SGI1-D, SGI1-K, SGI1-LK1) with deletions in backbone regions.
Performed complementation experiments using specific gene fragments (traHG-S010, S013-S014).

Main Results:

Complete SGI1 backbones (SGI1-I, SGI1-D) exhibited significant plasmid exclusion (40-100-fold).
Deletion of a 5793 bp region including traHG and S010 in SGI1-K/SGI1-LK1 abolished exclusion.
Complementation with traHG-S010 or S013-S014 fragments did not restore exclusion activity in SGI1-LK1.

Conclusions:

The SGI1 backbone, particularly the traHG-S010 region, plays a role in plasmid exclusion.
S010 is co-transcribed with traHG under an inducible promoter.
The precise genetic element(s) responsible for SGI1-mediated plasmid exclusion remain elusive despite identifying key deleted regions.