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

• Type of Material: Article
• Authors: Anton V. Bryksin, Emory University; Ichiro Matsumura, Emory University
• Language: English
• Date: 2010-10-08
• Publisher: Public Library of Science
• Publication Version: Final Published Version
• Copyright Statement: © 2010 Bryksin, Matsumura
• License: Creative Commons Attribution 4.0 International
• Final Published Version (URL): http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0013244
• Title of Journal or Parent Work: PLoS ONE
• ISSN: 1932-6203
• Volume: 5
• Issue: 10
• Start Page: 1
• End Page: 9
• Grant/Funding Information: The work was supported by the National Institutes of Health grants 1 R01 GM074264 and 1 R01 GM086824 to IM.
• Research Categories: Chemistry, Biochemistry

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.