Fig. 6. Mapping nuclear import signals in Dd-STATc. (A) Analysis of an N to C deletion series of Dd-STATc. A set of constructs, bearing the indicated N to C deletions, was constructed by PCR. The indicated regions were amplified and cloned immediately downstream of the Actin 15 promoter:GFP fusion present in GFP:STATc replacing the original Dd-STATc sequences (Fukuzawa et al., 2001). The constructs were transformed into cells and analysed for DIF inducibility as in Fig. 2B. (B) Identification of a nuclear import signal at the N terminus of Dd-STATc. Construct IMP:GFP contains the N-terminal-proximal 46 residues of Dd-STATc fused upstream of GFP. This construct contains a G418 resistance cassette and was transformed into Dictyostelium. Clones of expressing cells containing multiple copies of IMP:GFP were selected using G418. GFP accumulation in the nucleus was monitored in growing cells (left) and, in parallel, with an identical construct expressing GFP alone (right).

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