FIG 3 .

l-Arabinose inducible production of R. capsulatus CcoA in E. coli and R. capsulatus. (A) R. capsulatus ccoA was first cloned into the expression plasmid pBAD/Myc-His (pBK68) to provide an inducible and C-terminally Myc-His epitope-tagged ccoA construct. The plasmid pBK68 was then integrated into the broad-host-range plasmid pRK415 to yield pBK69, which is conjugally transferable into R. capsulatus. (B) Immunoblot analysis of R. capsulatus CcoA expressed in E. coli and in R. capsulatus. Membranes prepared from l-Ara induced E. coli (LMG194) and R. capsulatus ΔccoA (SE8) strains harboring appropriate plasmids expressing wild-type CcoA or mutant derivatives (as indicated), grown with or without 0.5% l-Ara, and probed with anti-Myc monoclonal antibodies.