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Plasmids

Plasmids

Plasmids are extrachromosomal DNA molecules found in bacteria, archaea, and some eukaryotic microbes like yeast. These small, circular DNA structures typically contain fewer than 30 genes, although some may exist linearly. Plasmids vary in their number within a cell, known as copy number. Single-copy plasmids are present in one copy per cell and multi-copy plasmids are present in multiple copies, reaching over 100 copies per cell.Plasmids usually replicate independently of the chromosomal DNA...

Mechanism of Conjugation

Mechanism of Conjugation

Bacterial conjugation is a mechanism of horizontal gene transfer that enables the exchange of genetic material between bacterial cells through direct contact. This process is facilitated by a donor cell carrying a conjugative plasmid, which encodes genes necessary for pilus formation, DNA replication, and transfer. The conjugative plasmid plays a central role in initiating and executing the transfer of genetic material.The tra region of the conjugative plasmid encodes proteins responsible for...

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Updated: Jun 23, 2026

Standardized Modular Assembly of Polycistronic Operons with Modular Cloning (MoClo) using the In-Cloning toolkit

Standardized Modular Assembly of Polycistronic Operons with Modular Cloning (MoClo) using the In-Cloning toolkit

Published on: September 2, 2025

A modular system for Clostridium shuttle plasmids.

John T Heap¹, Oliver J Pennington, Stephen T Cartman

¹BBSRC Sustainable BioEnergy Centre, School of Molecular Medical Sciences, Centre for Biomolecular Sciences, The University of Nottingham, University Park, Nottingham, NG7 2RD, UK. john.heap@nottingham.ac.uk

Journal of Microbiological Methods

|May 19, 2009

Summary

This summary is machine-generated.

Developing standardized modular shuttle plasmids for Clostridium research significantly advances genetic tools. This system enhances molecular biology for Clostridium species, crucial for medical and industrial applications.

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CRISPR-based Shuttle Cloning: A High-throughput Cloning Method

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Area of Science:

Microbiology
Molecular Biology
Genetics

Background:

The genus Clostridium holds significant medical and industrial importance, yet its fundamental biology is poorly understood compared to aerobic bacteria.
A key limitation is the lack of sophisticated genetic tools for clostridial molecular biologists.
Existing shuttle plasmid components for Clostridium-Escherichia coli are fragmented and underexplored.

Purpose of the Study:

To design and construct a standardized modular system for Clostridium-Escherichia coli shuttle plasmids.
To integrate and evaluate existing and novel genetic elements for improved clostridial genetic manipulation.
To provide a versatile platform for diverse applications in Clostridium research.

Main Methods:

Incorporation of established replicons and selectable markers from various plasmids.
Development and inclusion of a novel clostridial replicon.
Systematic comparison and characterization of individual shuttle plasmid components.

Main Results:

A standardized modular system for Clostridium-Escherichia coli shuttle plasmids was successfully designed and constructed.
Properties of individual components, including a novel replicon, were compared.
The system demonstrated extensive utility in laboratory settings for recombinant work.

Conclusions:

The developed modular system offers a standardized approach to shuttle plasmid construction for Clostridium.
This standardized system is expected to significantly benefit the Clostridium research community.
Researchers are encouraged to adopt and contribute to this evolving system for enhanced genetic studies.