Figure 5

The T-DNA insertion in CBP1 prevents production of the Cbp1 protein. Culture supernatants from the cbp1::T-DNA insertion (OSU8) lack the Cbp1 protein whereas culture supernatants from CBP1(+) yeast cells (WU15) show abundant production of Cbp1. Cell-free culture supernatants were prepared from late log/early stationary phase cultures of Histoplasma yeast and the major secreted proteins separated by electrophoresis. The Cbp1 protein runs as a 9-11 kD band. Positive identification of this band as Cbp1 was determined by loss of the 9-11 kD protein band from supernatants derived from a CBP1-RNAi strain (OSU38). A strain harboring a gfp-RNAi plasmid (OSU37) was used to show specific depletion of Cbp1 by CBP1-RNAi in OSU38. The secreted 20 kD protein produced by all strains was used to normalize supernatant loadings.