Fig. 1.

Construction of the primary vector Exp5(+). The XbaI and HindIII sites that precede the actin15 promoter in the existing Dictyostelium expression vector Exp4(+) were filled in and religated, which deleted both sites and created a novel unique NheI site. The small polylinker of Exp4(+) was subsequently exchanged with a larger multiple cloning site in order to accommodate dual TAP and YFP/GFP tags with at least three more restriction sites to insert the gene of choice.