The revolution in “genetic engineering” was predicated upon the presumption that any gene from any organism can be expressed in the laboratory bacterium, Escherichia coli. Three decades of subsequent frustration have clearly shown that E. coli is unable to properly fold most foreign proteins. To facilitate the heterologous expression of proteins in other bacteria, we have minimized the rolling circle replication origin of pWV01 and constructed a broad host range expression vector pBAV1K (Biobrick Accepting Vector). We have demonstrated the ability of this vector to replicate and produce reporter proteins in a variety of unrelated bacteria, including E. coli, Acinetobacter baylyi, Borrelia burgdorferi, Streptococcus pyogenes. Protein engineers can use this unique plasmid to try expressing their favorite genes in these widely divergent bacteria without any additional cutting or pasting of DNA.

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