FIG. 3.

Northern blot and RT-PCR analysis of cap1/cap1 mutants. CAP1 mRNA is absent in the cap1/cap1 strain and present at equivalent low levels in other strains during yeast growth (A) or germ tube induction (B). Total RNA (7 μg/lane), isolated as described in Materials and Methods, was separated in a formaldehyde agarose gel transferred to a nitrocellulose membrane and probed with radiolabeled PCR-1.2 to detect CAP1 mRNA and 18S rRNA as a control. The membrane was exposed to X-ray film for 7 days for detection of CAP1 mRNA and for 4 h for detection of 18S rRNA. (C) Amplification of 5′ (605 bp, 1 to 605) and 3′ (713 bp, 922 to 1634) portions of CAP1 mRNA using RT-PCR followed by Southern blotting using radiolabeled PCR-1.6 as probe. ACT1 mRNA (304 bp) was amplified as a positive control. Lanes 1 to 4, strains UnoPP-1, CAC1, CAC1-1A, and CACRE1.