Separation and Purification of GST-glycerol-3 -phosphate Dehydrogenase

摘要

In order to investigate the expression of glycerol-3-phosphate dehydrogenase by GCY1 gene in recombinant Saccharomyces cerevisiae,induction culture of the S.cerevisiaestrain was performed with SD-URA 2% galactose,3 × YP + 6% glucose,SC-URA 2% galactose,and SC-URA 2% galactose + 5% Na Cl glycerol-3-phosphate dehydrogenase,the cultured S.cerevisiaewas comminuted followed by full-automatic high-speed purification,and SDS-PAGE gel electrophoresis was performed for molecular weight of the GST fusion protein.The results showed that after shaking culture of the S.cerevisiae containing GCY1 at 25 ℃,the OD values of its 3 × YP + 6% glucose culture and SC-URA 2% galactose + 5% Na Cl culture were 8.75 and 7.35,respectively.It was shown by purification with a Profinia low-pressure liquid chromatograph that only the S.cerevisiae cultured in SC-URA 2% galactose + 5% Na Cl medium expressed glycerol-3-phosphate dehydrogenase,the molecular weight of which was detected as 65 ku by SDS-PAGE gel electrophoresis.