FIG 3.

Immunodetection of Lo18 in the O. oeni recombinant strains. Strains carrying pSIPSYN (Oosyn), pSINSYNAS18-UTR (OoAS18-UTR), or pSIPSYNAS18 (OoAS18) vectors were cultivated at 30°C in FT80m medium until the end of the exponential growth phase. Total cellular proteins were extracted immediately (30°C) or following a thermal treatment (30 min at 42°C). For each strain and each condition, 5 μg of protein was loaded on a PVDF membrane activated with methanol. After drying the membrane, the O. oeni protein Lo18 was detected by immunodetection with polyclonal antibodies directed against Lo18 and anti-rabbit antibody conjugated with peroxidase. Signal intensity was measured by Image Lab 4.1 (Bio-Rad) to assess relative quantities of the Lo18 protein in each spot.