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.
Center for Fundamental and Applied Molecular Evolution, Department of Biochemistry, Emory University, Atlanta, Georgia, United States of America.
Anton V
AV
Bryksin
Ichiro
I
Matsumura
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