Figure 5.

Analysis of C. parasitica Gα subunit CPG-1 mRNA and protein accumulation in wild-type strain EP155 and corresponding mutants cultured under identical growth conditions. (A) Northern hybridization analysis of cpg-1 transcript accumulation in wild-type strain EP155, cpg-1 disruptant Δcpg-1, cpgb-1 disruptant Δcpgb-1, bdmA, and bdm-1 disruptant Δbdm-1. The blot was probed with the entire cpg-1 cDNA clone. (B) Western blot analysis of CPG-1 levels in the same strains analyzed in A with affinity purified anti-CPG-1 antibody as described (36). Signal intensity of visualized CPG-1 on poly(vinylidene difluoride) membranes was quantified with National Institutes of Health image 1.61 in combination with a umax powerlook 2000 scanner. Purified recombinant CPG-1 (6-His fusion) was used as a standard.