Figure 2.

Effect of different CBS gene mutations on the growth of yeast cells. (A) Yeast stains cells were grown in selection media for 4 d at 28oC in the presence or absence cysteine. (B) Western blot analysis. Yeast cell lysates were prepared and examined for CBS expression by both denaturing and native PAGE gels followed by immunoblot. For the denaturing gel, modified alkaline protein extraction method was used using equal weights 0.03 g of cells and loading 20μl per lane. For the native-PAGE: 48μg of total protein was loaded per lane. CBS activity (underneath the native) was expressed in nmol/mg of total protein per h. (C) Quantitative measurement of CBS protein expression level by measuring the band intensity by ImageJ software using three different membranes. Data represented as mean± SD of three experiment each done in triplicate n=9. Statistical analysis for the effect between different strains was measured by two-way ANOVA followed by Bonferroni post-hoc test (*p<0.05).