The expression of a rice Cu/Zn superoxide dismutase （Cu/Zn-SOD） in Saccharomyces cerevisiae regulated by GAPDH promoter, involved in the inhibition of endogenous Glutathione （GSH） synthesis, and the competitive ex...
详细信息
The expression of a rice Cu/Zn superoxide dismutase （Cu/Zn-SOD） in Saccharomyces cerevisiae regulated by GAPDH promoter, involved in the inhibition of endogenous Glutathione （GSH） synthesis, and the competitive expression was detected by constructing the expression vector transferred Cu/Zn-SOD gene into wild-type S. cerevisiae. Transcription and expression of the Cu/Zn-SOD gene in S. cerevisiawere were confirmed by northern blot and SDS-PAGE, respectively, and activity of the Cu/Zn-SOD from crude extracts was enzymatically detected based on the effect of nitroblue tetrazolium （NBT） after running a native polyacrylamide gel. The GSH synthesis was also tested by DTNB （5, 5＇-Dithiobis （2-nitrobenzoic acid）） method. Results showed that GSH synthesis was evidently suppressed by the expression of Cu/Zn-SOD gene in both control and heat shock strains. It implied that the expression of the Cu/Zn-SOD gene in S. cerevisiae has more potential facility in response to oxidative exposure than that of endogenous GSH, although Cu/Zn-SOD and GSH were both contributed to the function of oxygen radical oxidoreduction.
暂无评论