Novel method for covalent fluorescent labeling of plasmid DNA that maintains structural integrity of the plasmid.
Bioconjug Chem
; 11(1): 51-5, 2000.
Article
in En
| MEDLINE
| ID: mdl-10639085
ABSTRACT
We have developed a chemical strategy for covalent coupling of fluorophores to plasmid DNA. A p-azido-tetrafluoro-benzyl-lissamine conjugate was synthesized and purified. This conjugate was used to covalently associate fluorescent molecules to plasmid DNA by photoactivation. In contrast to nick-translated plasmid DNA, plasmid-lissamine conjugates appeared on gel as supercoiled DNA. Reporter gene was expressed after transfection of the plasmid-lissamine conjugates in NIH 3T3 cells, although gene transfer efficiency was decreased by 60% as compared with unlabeled DNA. Intracellular traffic of plasmid-lissamine conjugates was studied in transfected cells. After cytoplasmic microinjection, fluorescent plasmid did not diffuse from the site of injection and appeared to be progressively degraded in the cytoplasm.
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Collection:
01-internacional
Database:
MEDLINE
Main subject:
Plasmids
/
DNA
/
Naphthoquinones
/
Fluorescent Dyes
/
Naphthalenesulfonates
Limits:
Animals
Language:
En
Journal:
Bioconjug Chem
Journal subject:
BIOQUIMICA
Year:
2000
Document type:
Article