Abstract
The nuclear localization signal (NLS) of the SV40 large T antigen efficiently induces nuclear targeting of proteins. We have developed a chemical strategy for covalent coupling of NLS peptides to plasmid DNA. A p-azido-tetrafluoro-benzyl-NLS peptide conjugate was synthesized. This conjugate was used to covalently associate NLS peptides to plasmid DNA by photoactivation. Reporter gene was expressed after transfection of the plasmid-NLS conjugates in NIH 3T3 cells. The conjugates interacted specifically with the NLS-receptor importin alpha, but plasmid-NLS conjugates were not detected in the nucleus, by fluorescence microscopy, after cytoplasmic microinjection.
Publication types
-
Research Support, Non-U.S. Gov't
MeSH terms
-
3T3 Cells
-
Animals
-
Chromatography, High Pressure Liquid
-
Genes, Reporter
-
Glutathione Transferase / genetics
-
Mice
-
Microinjections
-
Microscopy, Fluorescence
-
Nuclear Localization Signals* / genetics
-
Nuclear Localization Signals* / radiation effects
-
Nuclear Proteins / chemistry*
-
Nuclear Proteins / genetics
-
Peptides / chemistry
-
Peptides / radiation effects
-
Plasmids / chemistry*
-
Plasmids / genetics
-
Plasmids / radiation effects
-
Recombinant Fusion Proteins / genetics
-
Recombinant Fusion Proteins / pharmacology
-
Transfection
-
Ultraviolet Rays
-
alpha Karyopherins
Substances
-
Nuclear Localization Signals
-
Nuclear Proteins
-
Peptides
-
Recombinant Fusion Proteins
-
alpha Karyopherins
-
Glutathione Transferase