FIG. 2.

Binding of selected host proteins to p33 replication protein in vitro. MBP-tagged p33C and MBP (1 μg each) were separately immobilized on amylose beads, followed by incubation with cytosolic extracts prepared from yeast individually expressing the given GST-His₆-tagged host proteins. The bound host proteins were eluted from the beads and were analyzed by 10% SDS-PAGE and detected via anti-His₆ antibody. We tested a total of 17 host proteins out of the 58 host proteins identified via the protoarray for their abilities to bind to p33C. Protein bands of the expected sized are depicted with arrows. Molecular mass markers are shown on the left. Note that Hbs1p was expressed as a His₆-tagged protein from plasmid pYES.