1887

Abstract

The green fluorescent protein (GFP) of has been developed here as a reporter for gene expression and protein localization in When wild-type (wt) GFP was expressed in it was not possible to detect fluorescence or a translation product for the wt protein. Since this was probably due in part to the presence of the non-canonical CTG serine codon in the sequence, this codon was changed to the leucine codon TTG. cells expressing this construct contained GFP mRNA but were non-fluorescent and contained no detectable translation product. Hence a codon-optimized GFP gene was constructed in which all of the 239 amino acids are encoded by optimal codons for In this gene were also incorporated two previously identified mutations in the chromophore that increase GFP fluorescence. cells expressing this yeast-enhanced GFP gene (yEGFP3) are fluorescent and contain GFP protein. yEGFP3 can be used as a versatile reporter of gene expression in and and the optimized GFP described here should have broad applications in these and other fungal species.

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1997-02-01
2021-07-24
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