Gourbatsi, E. and Al-Fageeh, M.B. and Marchant, R.J. and Scott, S.J. and Underhill, M.F. and Smales, C.M. (2006) Noncovalently linked nuclear localization peptides for enhanced calcium phosphate transfection. Molecular Biotechnology, 33 (1). pp. 1-11. ISSN 1073-6085.
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Abstract
The generation of cell lines stably expressing recombinant material is a lengthy process and there has thus been much interest in the use of transient expression systems to rapidly produce recombinant material. To achieve this, the DNA of interest must be delivered into the nucleus of the target cell. The mechanisms by which this process occurs are poorly understood and the efficiency of various methods differs widely. Recently, nuclear localization signals (NLSs) have been investigated to target entry of DNA into the nucleus of mammalian cells. We have used NLSs from the SV40 and Tat antigens mixed with our model luciferase reporter gene plasmid for the transfection of Chinese hamster ovary (CHO) cells using calcium phosphate and FuGENE 6 transfection technology. The noncovalent complexation of NLSs with plasmid DNA before calcium phosphate-mediated transfection resulted in enhanced reporter gene expression with increasing ratios of NLS to plasmid until reaching a maximum. At higher ratios than maximum expression, the expression On the other hand, when using FuGene 6 expression reagent NLSs did not enhance reporter levels decreased. On the other hand, when using FuGene 6 expression. Cell cycle arrest in G(2)/M phase obliterated the effect of the NLS on reporter gene expression when using the calcium phosphate transfection method.
| Item Type: | Article |
|---|---|
| Uncontrolled keywords: | nuclear localization signal; transient transfection; calcium phosphate transfection; enhanced gene expression; luciferase; cell cycle arrest |
| Subjects: | Q Science |
| Divisions: | Faculties > Science Technology and Medical Studies > School of Biosciences |
| Depositing User: | Mark Smales |
| Date Deposited: | 03 Sep 2008 07:34 |
| Last Modified: | 28 Mar 2012 10:50 |
| Resource URI: | http://kar.kent.ac.uk/id/eprint/6228 (The current URI for this page, for reference purposes) |
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