A HUMANIZED GREEN FLUORESCENT PROTEIN CDNA ADAPTED FOR HIGH-LEVEL EXPRESSION IN MAMMALIAN-CELLS

We constructed gfp(h), a synthetic version of the jellyfish Aequorea v ictoria green fluorescent protein (gfp) cDNA that is adapted for high- level expression in mammalian cells, especially those of human origin. A total of 92 base substitutions were made in 88 codons in order to c hange the codon usage within the gfp10 coding sequence so that it was more appropriate for expression in mammalian cells, We also describe a series of versatile recombinant adeno-associated virus and adenovirus vectors for delivery and expression of genes into mammalian cells and , using these vectors, demonstrate the efficient transduction and expr ession of the gfp(h) gene in the human cell line 293 and also in vivo, within neurosensory cells of guinea pig eye. Cells infected with reco mbinant adeno-associated virus-GFP(H) can be readily sorted by fluores cence-activated cell sorting, suggesting that the newly designed gfp(h ) gene could he widely used as a reporter in many gene delivery techno logies, including human gene therapy.