When bacteria are used to make medicinally useful proteins by transformation, the protein of interest must be separated from all of the other cellular proteins. In this experiment, the unique fluorescent properties of GFP and BFP will be used as an assay during their purification from an E. coli extract. The column fractions containing GFP or BFP will be identified by fluorescence and then purified. As an optional activity, purified protein fractions can be separated by SDS polyacrylamide gel electrophoresis (SDS-PAGE) to estimate the purity and size of the GFP and BFP proteins.
Purification & Size Determination of Green & Blue Fluorescent Proteins
When bacteria are used to make medicinally useful proteins by transformation, the protein of interest must be separated from all of the other cellular proteins. In this experiment, the unique fluorescent properties of GFP and BFP will be used as an assay during their purification from an E. coli extract. The column fractions containing GFP or BFP will be identified by fluorescence and then purified. As an optional activity, purified protein fractions can be separated by SDS polyacrylamide gel electrophoresis (SDS-PAGE) to estimate the purity and size of the GFP and BFP proteins.
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