FIG. 2.

Southwestern blot analysis of extracts prepared from the yeast and mycelial phases of H. capsulatum strains. (A) Nuclear (Nucl) and cytoplasmic (Cyto) extracts (40 μg) prepared from the yeast and mycelial phases of the Downs and G217B strains were electrophoresed on SDS–10% polyacrylamide gels, fixed, and stained with Coomassie brilliant blue. (B) A second gel, electrophoresed in parallel under the same conditions, was transferred by electroblotting, blocked, and treated with 10⁵ cpm of the PCR-amplified target probe (395 bp of the 5′ regulatory region of yps 21:E-9 in G217B)/ml for 2 h. The blot was washed and autoradiographed to demonstrate a protein (designated p30M) of an apparent molecular mass of 30 kDa from the mycelial phase nuclear extracts of G217B which interacts with the ³²P-labelled DNA target in situ.