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Exhaustive in vivo labelling of plasmid DNA with BrdU for intracellular detection in non-viral transfection of mammalian cells

Title data

Jérôme, Valérie ; Heider, Andreas ; Schallon, Anja ; Freitag, Ruth:
Exhaustive in vivo labelling of plasmid DNA with BrdU for intracellular detection in non-viral transfection of mammalian cells.
In: Biotechnology Journal. Vol. 4 (October 2009) Issue 10 . - pp. 1479-1487.
ISSN 1860-7314
DOI: https://doi.org/10.1002/biot.200900053

Official URL: Volltext

Abstract in another language

The study of the non-viral gene delivery process at the molecular level, e.g. during the transfection of mammalian cells, is currently limited by the difficulties of specifically detecting the transfected plasmid DNA within the cells. Here we describe the in vivo production of 5-bromodeoxyuridine (BrdU)-labelled plasmid DNA by a thymine-requiring Escherichia coli strain leading to 92 +/- 15% BrdU incorporation while minimizing plasmid structure alteration. The labelled plasmid is produced on the milligram scale in a two-stage cultivation process. The relevance of this approach for plasmid DNA visualisation in the field of gene delivery is demonstrated by localising the BrdU-labelled plasmid DNA via immunodetection/fluorescence microscopy in CHO-K1 cells after electroporation with naked, BrdU-labelled plasmid DNA and after polyfection with polyethylenimine/BrdU-labelled plasmid complexes.

Further data

Item Type: Article in a journal
Refereed: Yes
Keywords: 5-Bromodeoxyuridine; DNA detection; Gene delivery; In vivo labelling; Transfection
Institutions of the University: Faculties > Faculty of Engineering Science
Faculties > Faculty of Engineering Science > Chair Process Biotechnology
Faculties > Faculty of Engineering Science > Chair Process Biotechnology > Chair Process Biotechnology - Univ.-Prof. Dr. Ruth Freitag
Faculties
Result of work at the UBT: Yes
DDC Subjects: 500 Science
500 Science > 500 Natural sciences
Date Deposited: 24 Feb 2016 13:22
Last Modified: 24 Feb 2016 13:22
URI: https://eref.uni-bayreuth.de/id/eprint/31031