Figure 7

Quantification of E2 protein levels in different congenic K. lactis strains. A) Dot blots performed in triplicates with 10, 25 and 50 μg of crude cell lysates from parent strain VAK367, the E2-producing strain VAK726; the Klgal80 mutant derivative of VAK726 (VAK746); and the two derivatives of VAK726 carrying two (VAK834) and five (VAK836) copies of the KlGAL4 gene, respectively. Cell lysates were applied to nitrocellulose membranes and incubated with E2-specific antibody WB-214 (PA2020; c.c.pro, Germany) and polyclonal secondary antibody (Dylight 488; Dianova). B) Relative fluorescence signal intensities were quantified as described in Material and Methods and are given relative to strain VAK726.