A "humanized" green fluorescent protein cDNA adapted for high-level expression in mammalian cells.
AUTOR(ES)
Zolotukhin, S
RESUMO
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 gfph 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.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=190401Documentos Relacionados
- High-level expression of human insulin receptor cDNA in mouse NIH 3T3 cells.
- Cloning and High-Level Expression of α-Galactosidase cDNA from Penicillium purpurogenum
- High-level expression of a heterologous protein in the milk of transgenic swine using the cDNA encoding human protein C.
- LCR/MEL: a versatile system for high-level expression of heterologous proteins in erythroid cells.
- Stuttering: high-level mistranslation in animal and bacterial cells.