Figure 3

Construction of a M. mycoides with re-structured genome. (A) BssHII restriction enzyme map of the M. mycoides genome with an inversion of the 100 kb synthetic DNA. (B) Transplantation colonies from wild type and two semi-synthetic genomes. Transplant colonies by genomes from yeast clones (from left to right): wild type genome (WT), the 100 kb-forward insertion genome (For) and the 100 kb-inverted insertion genome (Inv). (C) CHEF electrophoresis analysis of BssHII-digested M. mycoides genomes. M. mycoides genomes purified from bacterial transplants were digested with BssHII, lane 1: WT genome, lane 2: Inv genome (clone 1), and lane 3: Inv genome (clone 2). Two DNA fragments (212 and 204 kb) co-migrated shown in lane 2 and 3, indicated by yellow arrows. M: 50 kb lambda DNA ladder.