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Directed protein evolution often employs random mutagenesis to spawn mutant sequences of genes of interest that can be expressed and screened to identify new proteins with altered characteristics.
John N. Davis and Anthony N. van den Pol of Yale University School of Medicine, New Haven, have developed a novel methodology for this task, that uses an RNA virus with a high normal mutation rate due to lack of polymerase proofreading, into which the gene of interest is inserted, says van den Pol. "The virus not only mutates the gene, but it also expresses the mutant protein coded by the gene in mammalian cells that the virus infects." The big advantage: "in a single day, the virus can generate hundreds of thousands of progeny virus, each with a high probability of having a different single mutation. This is particularly useful for generating new fluorescent proteins, as mutations can shift the fluorescence to different colors. If a simple way of detecting the mutated protein were available, this approach could be used for a wide variety of genes." This methodology could be used to look for fluorescence proteins that change properties under different physiological conditions, so that, for example, a mutant fluorescent protein sensitive to shifts in calcium or pH could be detected by time-lapse imaging of virally infected cells.
(J. N. Davis and A. N. van den Pol. 2010. Viral mutagenesis as a means for generating novel proteins. J. Virol. 84:1625-1630.)
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