Figure 6

Investigation of amount of DNA and outgrowth duration on the electrotransformation of C. pasteurianum . A. Effect of plasmid DNA amount on total number of transformants and electrotransformation efficiency of C. pasteurianum. Separately, 0, 0.25, 0.5, 1.0, 2.5, and 5.0 μg of pMTL85141 were added to electrocompetent cells of C. pasteurianum and electroporated. Total number of thiamphenicol-resistant transformants (light shading) and electrotransformation efficiency (dark shading) were quantified. Pulse duration was unaffected between samples. Zero μg pMTL85141 failed to generate thiamphenicol-resistant transformants. B. Effect of post-electroporation incubation time. Cells were electroporated, transferred to 10 ml outgrowth medium containing 0.2 M sucrose, and incubated for 0, 2, 4, 6, or 16 hours prior to selective plating.