rational design of a plasmid origin that replicates efficiently in both gram-positive and gram-negative bacteria设计合理的质粒的起源在革兰氏阳性和革兰氏阴性细菌有效复制.pdfVIP

Rational Design of a Plasmid Origin That Replicates Efficiently in Both Gram-Positive and Gram-Negative Bacteria Anton V. Bryksin, Ichiro Matsumura* Center for Fundamental and Applied Molecular Evolution, Department of Biochemistry, Emory University, Atlanta, Georgia, United States of America 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. Citation: Bryksin AV, Matsumura I (2010) Rational Design of a Plasmid Origin That Replicates Efficiently in Both Gram-Positive and Gram-Negative Bacteria. PLoS ONE 5(10): e13244. doi:10.1371/journal.pone.0013244 Editor: Igor Mokrousov, St. Petersburg Pasteur Institute, Russian Federation Received July 25, 2010; Accepted September 8, 2010; Published October 8, 2010 Copyright: 2010 Bryksin, Matsumura. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Funding: The work was supported by the National Institutes of Health grants 1 R01 GM074264 and 1 R01 GM086824 to IM. The funder had no role in study design, data collection and analysis, deci