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Protein Overexpression
Exploring Mutant Proteins
Screening for Mutants
Your trick is simple. You fuse the gene encoding UDP glucose dehydrogenase to that encoding green fluorescent protein (GFP). If the enzyme interacts to produce a precipitate, then GFP will get pulled into the precipitate as well and fail to fluoresce. A mutant enzyme that fails to precipitate will leave the tethered GFP free as well, giving rise to a highly fluorescent colony.
That's the theory. In fact what you get are a few rare colonies that show moderately more fluorescence than the unmutated fusion. These rare colonies show less precipitate, but there's still a lot. You're going in the right direction, but you clearly need to go further -- further than random mutagenesis can take you.
Photo courtesy of Clare Chemical Research
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