Rational design of a plasmid origin that replicates efficiently in both gram-positive and gram-negative bacteria.
BACKGROUND
Most plasmids replicate only within a particular genus or family.


RESULTS
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
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.
Bryksin
Anton V
AV
Center for Fundamental and Applied Molecular Evolution, Department of Biochemistry, Emory University, Atlanta, Georgia, United States of America.
Matsumura
Ichiro
I
D001483:Base Sequence; D017931:DNA Primers; D004261:DNA Replication; D006090:Gram-Negative Bacteria / Q000145:classification / Q000235:genetics*; D006094:Gram-Positive Bacteria / Q000145:classification / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics; D010802:Phylogeny; D010957:Plasmids; D016133:Polymerase Chain Reaction
D017931:DNA Primers; D049452:Green Fluorescent Proteins