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|Monomer||Discosoma sp.||26.7 kDa||-|
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The mCherry-K92N/K138C/K139R/N196D variant, with just four of the six mCherry2 mutations, permitted a relative faster E. coli growth rate than EGFP, suggesting that this variant (designated as mCherry1.5) had even lower cytotoxicity than EGFP and mCherry2... Unfortunately, both mCherry1.5 and mCherry itself showed similar patterns of protein mislocalization in these fusions, as compared to identical mEGFP fusions, indicating that the low bacterial cytotoxicity of an RFP does not necessarily correlate with more faithful fusion protein localization. Based on this result and our cumulative experience with engineering RFPs, we suggest that ameliorating the mislocalization of RFPs may ultimately require a high-throughput image-based screen of mammalian cells expressing a library of RFP variants in the context of a fusion that tends to mislocalize.