Figure 6.—

Identification and characterization of csp-1. (A) Schematic of gene replacement at the endogenous locus. Blue arrow, NCU02713 ORF; blue line, left flank; red line, right flank; gray line and arrow, hygromycin-resistance cassette containing hph ORF; blue and yellow bars, WT and C139Y allele, respectively, at 6.194611. The resulting strains are csp-^1WT∷hph (pc^WT) and csp-1^C139Y∷hph (pc^mut), which differ only at the Cys139 locus of the NCU02713 ORF. (B) Phenocopying of csp-1 by NCU02713^KO and pc^mut but not pc^WT. Strains, including parental controls, were grown for 3 days at 30 LL on minimal medium and tapped sharply (top) or had their conidia transferred into water (middle). (Bottom) Genotype at the csp-1 locus. The C139Y mutation creates an additional RsaI restriction site (asterisk). (C) Representative Northern blots of csp-1 mRNA over a 48-hr time course, in WT (top) and ras-1^bd (bottom) background. Both sets were performed on the same membrane and with identical exposure times.