FIG. 5.

GST pull-down assay to probe the interaction between RYSV P3 and the N protein. Glutathione Sepharose 4B beads were loaded with purified GST-N fusion protein and then incubated with ³⁵S-labeled P3 protein. The bound protein was eluted and resolved on SDS-12% PAGE and autoradiographed (lane 2). GST-loaded beads (lane 3) and unloaded beads (lane 1) were used as controls, respectively. Lane 4, in vitro transcription and translation products using pGEM-P3 as a template. Black arrowhead, ³⁵S-labeled P3 protein; hollow arrowhead, a ³⁵S-labeled in vitro translation product that probably resulted from an internal ATG of the ORF3 sequence.