FIG. 1.

Yeast two-hybrid analysis of the interactions of HSV-1 tegument protein pUL48. (A) Target pUL48N (amino acids 1 to 411) was screened against a number of Bait fusion constructs using the LexA yeast two-hybrid assay. Summary of activity values obtained from a liquid β-galactosidase assay. The activity was calculated from the following equation: β-galactosidase activity = 1,000 × A₄₂₀/(t × V × OD₆₆₀), where t = time (in minutes) of incubation, V is the volume of cells (ml) used in the assay, and OD₆₀₀ is the optical density at 600 nm. The given activity values are the averages of measurements from at least three separate colonies. Positive control corresponds to Bait pUL11 plus Target pUL16. (B) Immunoblot with monoclonal anti-LexA showing expression of Bait fusion constructs in yeast. (C) Immunoblot with monoclonal anti-HA showing expression of Target fusion constructs in yeast. For both panels B and C, samples (from left to right) consisted of yeast cotransformed with the combination of Bait pUL11/Target pUL16, Bait pUL19UD/Target pUL48N, or Bait pUL38N/Target pUL48N. For determination of protein expression, samples were subjected to protein extraction before separation by SDS-PAGE (12%). The position of each fusion protein is indicated (*). pUL19UD (amino acids 451 to 1054), pUL36N (amino acids 1 to 767), and pUL38N (amino acids 1 to 90).