Figure 3

Suppression of Ca²⁺ sensitivity of the K667 yeast mutant by OsCCX2.

The yeast K667 triple mutant (∆pmc1∆vcx1∆cnb1) is unable to grow in the presence of higher CaCl₂ because low affinity Ca²⁺ transport is compromised in the mutant. The galactose inducible yeast expression vector, pYES2, was used to complement K667 with OsCCX2. The growth of pYES2/K667 (empty vector control) was compromised at higher CaCl₂ concentration either in presence of glucose or galactose in the medium. Yeast strain transformed with either vector or OsCCX2 were grown in SC-Ura media overnight at 30 °C and diluted to 0.5 OD₆₀₀ followed by 10 times serial dilutions of 0.5 OD₆₀₀ were dotted on SC-URA + Gal with different CaCl₂ concentrations. The plates were incubated at 30 °C for 3 days. A K667 yeast mutant expressing OsCCX2 could grow in presence of excess CaCl₂ while growth of empty vector expressed mutant was severely compromised.