A "Humanized" green fluorescent protein cDNA adapted for high-level expression in mammalian cells

Sergei Zolotukhin, Mark Potter, William W. Hauswirth, John Guy, Nicholas Muzyczka

Research output: Contribution to journalArticle

535 Citations (Scopus)

Abstract

We constructed gfph, a synthetic version of the jellyfish Aequorea victoria 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 change 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 expression of the gfp, gene in the human cell line 293 and also in vivo, within neurosensory cells of guinea pig eye. Cells infected with recombinant adeno-associated virus-GFPH can be readily sorted by fluorescence-activated cell sorting, suggesting that the newly designed gfph gene could be widely used as a reporter in many gene delivery technologies, including human gene therapy.

Original languageEnglish
Pages (from-to)4646-4654
Number of pages9
JournalJournal of Virology
Volume70
Issue number7
StatePublished - Dec 1 1996
Externally publishedYes

Fingerprint

Green Fluorescent Proteins
green fluorescent protein
Complementary DNA
Dependovirus
gene transfer
codons
Codon
cells
Aequorea victoria
Genes
gene therapy
human cell lines
Scyphozoa
Adenoviridae
Genetic Therapy
guinea pigs
flow cytometry
Flow Cytometry
Guinea Pigs
genes

ASJC Scopus subject areas

  • Immunology

Cite this

Zolotukhin, S., Potter, M., Hauswirth, W. W., Guy, J., & Muzyczka, N. (1996). A "Humanized" green fluorescent protein cDNA adapted for high-level expression in mammalian cells. Journal of Virology, 70(7), 4646-4654.

A "Humanized" green fluorescent protein cDNA adapted for high-level expression in mammalian cells. / Zolotukhin, Sergei; Potter, Mark; Hauswirth, William W.; Guy, John; Muzyczka, Nicholas.

In: Journal of Virology, Vol. 70, No. 7, 01.12.1996, p. 4646-4654.

Research output: Contribution to journalArticle

Zolotukhin, S, Potter, M, Hauswirth, WW, Guy, J & Muzyczka, N 1996, 'A "Humanized" green fluorescent protein cDNA adapted for high-level expression in mammalian cells', Journal of Virology, vol. 70, no. 7, pp. 4646-4654.
Zolotukhin, Sergei ; Potter, Mark ; Hauswirth, William W. ; Guy, John ; Muzyczka, Nicholas. / A "Humanized" green fluorescent protein cDNA adapted for high-level expression in mammalian cells. In: Journal of Virology. 1996 ; Vol. 70, No. 7. pp. 4646-4654.
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