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- Title
Separation and Purification of GST-glycerol-3 -phosphate Dehydrogenase.
- Authors
Hongmei ZHAO; Shihai LI; Yasuo Shihai LI; Yasuo WATANABE
- Abstract
In order to investigate the expression of glycerol-3 -phosphate dehydrogenase by GCY1 gene in recombinant Sacharomyces cerevisiae, induction culture of the S. cerevisiaestrain was performed with SD-URA 2% galactose, 3 X YP + 6% glucose, SC-URA 2% galactose, and SC-URA 2% galactose + 5% NaCl glycerol-3-phosphate dehydrogenase, the cultured S. cerevisiae was comminuted foUowed 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 °C, the OD values of its 3 X YP + 6% glucose culture and SC-URA 2% galactose + 5% NaCl culture were 8.7 5 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% NaCl medium expressed glycerol--phosphate dehydrogenase, the molecular weight of which was detected as 65 ku by SDS-PAGE gel electrophoresis.
- Subjects
GLYCEROLPHOSPHATE dehydrogenase; MOLECULAR weights; GENE expression; GEL electrophoresis; GALACTOSE
- Publication
Agricultural Biotechnology (2164-4993), 2016, Vol 5, Issue 5, p44
- ISSN
2164-4993
- Publication type
Article