FIG. 3.

Assay of HIV PR and B3 variant autoproteolysis in E. coli. (A and B) DHT1 cells were transformed with plasmids expressing AC-N22-PR-C29 (section 1), AC-N7-PR-C7 (section 2), AC-N22-B3-C29 (section 3), AC-N7-B3-C7 (section 4), ACp5 (section 5), or T25 (section 6) and plated on MacConkey-maltose plus kanamycin (A) or on the same medium supplemented with 100 μM saquinavir (B) and grown for 48 h at 30°C. (C) Recombinant ACs (ACp5, apparent molecular mass of 45 kDa; ACPR or ACB3, apparent molecular mass of 56 kDa), in DHT1 cells expressing the indicated proteins, were detected by Western blot analysis as described in Materials and Methods. β-Galactosidase activities and cAMP levels in the corresponding samples were measured as described in Materials and Methods.