at Google ScholarTitle 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
(2009)
Issue 10
.
- pp. 1479-1487.
ISSN 1860-7314
DOI: https://doi.org/10.1002/biot.200900053
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: | 26 Apr 2022 11:19 |
| URI: | https://eref.uni-bayreuth.de/id/eprint/31031 |