Gold Medal Criterion #2

pSB1C3 transformation into Enterobacter ludwigii FCP2-01

Our team used the plasmid backbone pSB1C3 in the organism Enterobacter ludwigiiFCP2-01 (BBa_K2633006), and provided the first evidence that an Enterobacter species can be transformed stably with this organism. The clade which our chassis comes from is important medically, agriculturally, and ecologically (Hoffmann and Roggenkamp 2003), and this provides potential to apply this part and many parts in the Registry of Standard Biological Parts to organisms in this group.

Our improvement upon pSB1C3 is by the composite part BBa_K2633000. A restriction illegal version of this part was used to transform Enterobacter ludwigii FCP2-01, as seen in Figure 1. The transformed strain was then used to inoculate Brachypodium distachyon by soaking seeds in bacterial culture, and the plant were grown for 5 weeks with images taken at 1 week intervals. Example of FCP2-01 expressing acdS-GFP are seen in Figure 2.

Figure 1. A) FCP2-01 transformed with GFP (left) vs wild-type (right) under blue light with orange filter. B) Transformed colonies on an agar plate.
Figure 2. A) Transformed FCP2-01 forming a macrocolony in root cortex of Brachypodium distachyon, viewed 3 weeks post-inoculation (WPI). B) Transformed FCP2-01 growing on root hairs and epidermis 4 WPI.