Figure 6

Tca2 pol protein expression is directed by LTR promoter activity in C. albicans. The pol ORF of a genomic copy of C. albicans Tca2 was tagged at its 3' end immediately before the pol stop codon with Protein A coding sequence. Panel A; proteins from the untransformed host strain (CAI-4) and a pol-Protein A integrant (ProA-Int) were separated (SDS-PAGE), western-blotted, and blots probed with anti-Protein A antibody. Arrowhead indicates the migration position of a 76 kDa protein induced under heat shock conditions, present in the integrant but not the parental strain CAI-4. Results are typical of at least three experiments involving independent Protein A integrant isolates. Panel B; a replicate gel was Coomassie Blue-stained to show equivalent lane loadings. Panel C; the anti-Protein A antibody specificity was confirmed by probing a control blot of untransformed CAI-4 lysate and that from an S. cerevisiae strain expressing TAP-tagged eIF4A translation factor [59]. The 65 kDa protein detected by the antibody (indicated by arrowhead) represents eIF4A (44.6 kDa) fused to the TAP tag (20 kDa), the latter containing Protein A.