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Author Notes:

Correspondence: Ichiro Matsumura, Email: imatsum@emory.edu

Authors' Contributions: Conceived and designed the experiments: AVB

Performed the experiments: AVB

Analyzed the data: AVB and IM

Wrote the paper: AVB and IM

Acknowledgments: We thank Felipe Cabello, Julia Bugrysheva, Dorothea Zahner, Shana Topp, David S. Weiss, Connie Arthur and Justin Gallivan for their guidance, and for their invaluable assistance with bacterial species or techniques new to us.

Disclosures: The authors have declared that no competing interests exist.

The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Subject:

Research Funding:

The work was supported by the National Institutes of Health grants 1 R01 GM074264 and 1 R01 GM086824 to IM.

Rational Design of a Plasmid Origin That Replicates Efficiently in Both Gram-Positive and Gram-Negative Bacteria

Tools:

Journal Title:

PLoS ONE

Volume:

Volume 5, Number 10

Publisher:

, Pages 1-9

Type of Work:

Article | Final Publisher PDF

Abstract:

Background: Most plasmids replicate only within a particular genus or family. Methodology/Principal Findings: Here we describe an engineered high copy number expression vector, pBAV1K-T5, that produces varying quantities of active reporter proteins in Escherichia coli, Acinetobacter baylyi ADP1, Agrobacterium tumefaciens, (all Gram-negative), Streptococcus pneumoniae, Leifsonia shinshuensis, Peanibacillus sp. S18-36 and Bacillus subtilis (Gram-positive). Conclusions/Significance: Our results demonstrate the efficiency of pBAV1K-T5 replication in different bacterial species, thereby facilitating the study of proteins that don't fold well in E. coli and pathogens not amenable to existing genetic tools.

Copyright information:

© 2010 Bryksin, Matsumura

This is an Open Access work distributed under the terms of the Creative Commons Attribution 4.0 International License (https://creativecommons.org/licenses/by/4.0/).
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