Yeast Experiments
From 2007.igem.org
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LucAssay result : | LucAssay result : | ||
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2XORE promoter have a minimal basal activity without oleate and we can see a proportionally increment when increasing oleate concentrations | 2XORE promoter have a minimal basal activity without oleate and we can see a proportionally increment when increasing oleate concentrations | ||
Revision as of 10:00, 10 September 2007
For luciferase assays and the preparation of protein extracts, transformants were grown in rich medium with 1.5% raffinose, 1.5% glycerol, and 1% ethanol as carbon sources. Induction was done with different concentrations of oleate overnight.Cells were lysed in 100 mM potassium phosphate pH 7.8, 1 mM phenylmethylsulfonyl fluoride, 1 mM dithiothreitol using glass beads (diameter 0.45 mm). Cell debris were removed by centrifugation at 15 000 x g at 4°C for 20 min.Luciferase activities are expressed in relative light units/pg protein and protein concentrations were determined by the method of Bradford using bovine serum albumin as a standard.
LucAssay result :
2XORE promoter have a minimal basal activity without oleate and we can see a proportionally increment when increasing oleate concentrations
