Figure 5.

Yeast two-hybrid assay. (a) Self-interaction of LmjMCA via its C-terminal domain. Interactions of the LmjMCA full-length form (FL), the N-terminal domain (NT), the catalytic domain (cd) and the C-terminal domain (CT) have been tested against each other by cloning them into the bait and/or the prey vector for yeast two-hybrid assay. The positive interactions (+) seemed mainly owing to the C-terminal domain of LmjMCA. (b) Number of positive colonies growing in a medium without leucine, tryptophan, histidine and adenine and with X-α-Gal, co-transfected with different GAL4-AD protein libraries (logarithmic promastigotes, stationary promastigotes, axenic amastigotes and intracellular amastigotes) and either the inactive full-length LmjMCA (147/202LmjMCA, in blue) or the inactive catalytic domain of LmjMCA (202 cd-LmjMCA, in green). A greater number of positive colonies was obtained with the full-length than with the catalytic domain