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Bacterial Transformation

Bacterial Transformation

In 1928, bacteriologist Frederick Griffith worked on a vaccine for pneumonia, which is caused by Streptococcus pneumoniae bacteria. Griffith studied two pneumonia strains in mice: one pathogenic and one non-pathogenic. Only the pathogenic strain killed host mice.
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Controllable Ion Channel Expression through Inducible Transient Transfection

Controllable Ion Channel Expression through Inducible Transient Transfection

Published on: February 17, 2017

High-throughput cloning and expression in recalcitrant bacteria.

Eric R Geertsma¹, Bert Poolman

¹Department of Biochemistry, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Nijenborgh 4, 9747 AG, Groningen, The Netherlands.

|July 24, 2007

Summary

This summary is machine-generated.

We created a new high-throughput cloning method for difficult bacteria. This technique uses vector-backbone exchange (VBEx) for rapid plasmid construction and efficient transformation, broadening genetic engineering capabilities.

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Area of Science:

Molecular Biology
Microbiology
Synthetic Biology

Background:

Conventional DNA manipulation methods face challenges in bacteria with limited genetic tractability.
Efficient cloning is crucial for genetic engineering and understanding microbial functions.

Purpose of the Study:

To develop a versatile, high-throughput cloning strategy applicable to bacteria that are difficult to engineer.
To establish a rapid and efficient method for generating organism-specific plasmids.

Main Methods:

A novel generic method combining ligation-independent cloning in an intermediary Escherichia coli vector.
Rapid conversion of the intermediary vector into an organism-specific plasmid via vector-backbone exchange (VBEx).

Main Results:

Demonstrated proof of principle for VBEx in Lactococcus lactis.
The method facilitates high-throughput cloning for bacteria with challenging DNA manipulations.

Conclusions:

The developed VBEx method offers a broadly adaptable platform for high-throughput cloning across diverse bacterial species.
This technique enhances the ease and efficiency of genetic engineering in previously recalcitrant organisms.