Yeast Experiments

From 2007.igem.org

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LucAssay result :
LucAssay result :
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                 [[Image:LucGraph.jpg]]
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                 [[Image:Luciferase1.jpg]]
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 :

                Luciferase1.jpg

2XORE promoter have a minimal basal activity without oleate and we can see a proportionally increment when increasing oleate concentrations