Fig. 2.
Structure determination of the predicted gene CND04970 and growth defect of a C. neoformans ggt1Δ mutant. (A) Gene models of the locus encoding Ggt1. Coding regions are depicted as thick black bars, introns by thin black lines and untranslated regions by thick gray bars. Model I shows the CNAG01385 annotation from the Broad Institute. Model II shows the annotation suggested by homology, existing EST evidence and preliminary polymerase chain reaction (PCR) studies (see text). Model III shows our revised annotation, as confirmed by PCR studies (not shown) and short read assemblies of cDNA. The contigs derived from RNA-Seq data are depicted below the gene models as thin black bars with intron-bridging indicated by grey peaks (RNA-Seq). Importantly, the predicted galactosyltransferase domain lies in the region that is common to all three models. (B) Growth defect and complementation of a C. neoformans ggt1Δ mutant. Wild-type and ggt1Δ mutant strains carrying empty vector, episomal GGT1short or GGT1long were spotted in serial 10-fold dilutions on YPD plates and incubated at the indicated temperatures for 3 days. Two independent transformants are shown for the plasmid containing the long form of the gene.