Map the interaction domains between pUL85 and pUL23. A) Schematic illustration of pUL85 structures and indicated region truncated. The cDNA fragments encoding region 1-111aa, 104-202aa and 196-307aa of pUL85 were obtained by PCR and cloned into vector pGADT7 and pGEX4T-1, respectively, for yeast two-hybrid assay and GST Pull-down assay; B) Mapping the pUL85 binding domain to pUL23 by yeast two hybrid. Yeasts transformants were streaked on SD/-Trp/-Leu and SD/-Trp/-Leu/-His/-Ade selective plates and cultured (Left). Yeast colonies were analyzed for expression of the reporter gene β-galactosidase by filter lift assays (Right); C) Mapping the pUL85 binding domain to pUL23 by GST Pull-down. GST protein or GST-fusion proteins of pUL85-N / M / C bound to glutathion-Sepharose bead were incubated with lysates prepared from cos-7 cells transfected with pcDNA3.1(+)-UL23-Flag.Bound proteins were analyzed by western blot (WB) with anti-Flag Antibody