Figure 4
Functional assay of CsHMA3 of the wild type and mutants in yeast. Confirmation of Cd and Pb sensitivity of Δycf1, and Zn sensitivity of Δzrc1 mutant strain compared with those of wild-type (BY4741) yeast strain (A). Cell growth of the Δycf1 or Δzrc1 mutant strain was transformed with empty vector pYES2, CsHMA3 (B). Yeast cells grown in the log phase were diluted to an OD600 of 1, 10-1, 10-2, 10-3, and 10-4, and 5 μL of each dilution were spotted onto the 1/2 YPD (A) or LPM medium containing galactose (B) without or with added CdSO4, PbNO3, and ZnSO4. The plates were incubated at 30°C for four days before photography. The yeast cells were grown in the presence of 25 μM Cd, 10 μM Pb, and 5 mM Zn. The photographs shown are representative results of independent experiments that were performed three times with reproducible results.