FIG. 5.

Restriction maps of pDA5 and its derivatives. Plasmid pDA5 carries a ubiE::ccmC fusion in frame spanning the internal EcoRI site joining the 1.6- and 1.4-kb fragments, which are cloned separately to yield pCS1534 and pCS1535, respectively. Plasmids p2hel-404 and pCS1532 carry ccmABCDG and ccmCDG, respectively. pCS1542 and pCS1539 are derivatives of pCS1532 with a ccmC::spe insertion polar on the downstream genes ccmDG and a frameshift mutation in ccmC (ccmΔCB), respectively. pCS1541, pCS1527 and pCS1540 are derivatives of pCHB500 and express ccmDG, ccmD and ccmG, respectively, via the cycA promoter. The ability of these plasmids to complement the CcmI-null mutant MT-SRP1.r1, the CcmD-null mutant SB1003 ΔhelD, the CcmG-null mutant SB1003 ΔhelX, and the CcmC-null mutant TCK2 for Ps⁺ growth on enriched medium is shown on the right. Curved arrows refer to various promoters mediating different levels of expression of downstream genes, as follows. Black arrow, E. coli lacZ promoter; dark gray arrow, ubiE promoter; light gray arrow, promoter downstream of the ccmB and upstream of the EcoRI site in ccmC; white arrow with black border, cycA promoter; gray arrow with black border, promoter downstream of the BamHI site in ccmC and upstream of ccmD). X, K, E, and B refer to cleavage sites of the restriction enzymes XbaI, KpnI, EcoRI, and BamHI, respectively.