Rational design of a plasmid origin that replicates efficiently in both gram-positive and gram-negative bacteria.

PLoS One · 2010

basic_science · Level V

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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.