Fig. 9. Expression of recombinant 185/333 proteins. (A) Expression in E. coli. Western blot analysis of recombinant 185/333 protein, Sp0032 (GenBank acc. no. DQ183168; E1 pattern) expressed in E. coli (M15 strain). Bacteria were induced with 1 mM IPTG for 4 hours at 25°C prior to collection. Lane 1, pelleted E. coli without 1 mM IPTG induction. Lane 2, pelleted E. coli with 1 mM IPTG induction. Lane 3, pelleted E. coli with 1 mM IPTG and analyzed with pre-immune serum. Pelleted cells were lysed and separated into pellet and supernatant. Lane 4, sonicated pellet. Lane 5, sonicated supernatant. Standards are shown to the left. (B) Expression in insect cells. Drosophila S2 cells were transfected with Sp0296, a cDNA of the E2 pattern (GenBank acc. no. DQ183121) (see Terwilliger et al., 2006) and induced with 100 mM CuSO₄. Recombinant 185/333 proteins were secreted into the culture medium, which was collected and analyzed by western blot with the anti-185/333 antisera. The expected monomer of 31.5 kDa (without post-translational modifications), putative dimers of 65 kDa, and multimers of 150 kDa (lane 2 only) were detected. Lane 1, 1 day after induction. Lane 2, 2 days after induction. Size standards are shown to the left.