Skip to main content
. 2004 Feb;78(3):1181–1194. doi: 10.1128/JVI.78.3.1181-1194.2004

FIG. 3.

FIG. 3.

Mapping of the IE62 binding site in IE63 protein. The IE62 pull-down assay was performed with IE63-MBP fusion proteins that had deletions of the conserved amino acid pairs in region 2. The ORF63 mutations were made in pMAL-c2X and expressed in E. coli (BL21-AI) as fusion proteins at the MBP C terminus. The recombinant proteins were affinity purified with amylose resin (New England Biolabs, Beverly, Mass.) and incubated with recombinant IE62. IE62 that bound to the IE63 recombinant proteins was detected with polyclonal rabbit anti-ORF62 antibody. IE62 binding is shown to intact IE63 (lane 1), the IE63 mutants ΔW53/E54 (lane 3), ΔF68/L69 (lane 5), ΔR86/R87 (lane 6), ΔM95/G96 (lane 7), ΔW107/E108 (lane 8), ΔL111/Q112 (lane 9), ΔL121/R122 (lane 10), and 5′Phos− (lane 11). No IE62 was recovered in the pull-down assay done with MBP alone (lane 2) or with the IE63-MBP fusion protein ΔR59/L60 (lane 4).