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Biochem Biophys Res Commun. 1999 Sep 7;262(3):638-42.

Targeting of green fluorescent protein expression to the cell surface.

Author information

  • 1Center for Molecular Imaging Research, Massachusetts General Hospital, Building 149, 13th Street, Charlestown, Massachusetts, 02129, USA.

Abstract

We have previously reported on GPI-anchored fusion proteins that bind radioactive isotopes. We targeted their expression to the cell surface to obtain a marker protein detectable by nuclear and optical imaging (1, 2). Here we suggest a novel approach for targeting a model protein (GFP) to the exoplasmic surface of the plasma membrane. An expression vector (pcPEP-GFP) was constructed containing GFP cDNA fused with the fragment encoding the N-terminal cytoplasmic domain and signal peptide/membrane anchoring domain of the rabbit neutral endopeptidase (PEP-GFP). Flow cytometry showed green fluorescence in 45% of cells transfected with GFP and in 34% of cells transfected with PEP-GFP (24 h after transfection). Fluorescence microscopy of fixed cells stained with rhodaminated anti-GFP antibodies showed positive reaction only in the case of PEP-GFP-transfected cells indicating cell-surface expression. The PEP-GFP fusion protein was identified as a component of the light microsomal and Golgi fractions by immunoblotting.

Copyright 1999 Academic Press.

PMID:
10471377
[PubMed - indexed for MEDLINE]
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