Comparison of bovine leukemia virus (BLV) and CMV promoter-driven reporter gene expression in BLV-infected and non-infec
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BioMed Central
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Comparison of bovine leukemia virus (BLV) and CMV promoter-driven reporter gene expression in BLV-infected and non-infected cells Jerome S Harms*1, Kurt A Eakle2, Lillian S Kuo1, Robert D Bremel3 and Gary A Splitter1 Address: 1Department of Animal Health and Biomedical Sciences, University of Wisconsin-Madison, Madison, WI 53706-1581, USA, 2GALA Biotech, 8137 Forsythia Street, Middleton, WI 53562, USA and 3 IoGenetics LLC, 3591 Anderson St., Suite 218, Madison, WI 53704, USA Email: Jerome S Harms* - [email protected]; Kurt A Eakle - [email protected]; Lillian S Kuo - [email protected]; Robert D Bremel - [email protected]; Gary A Splitter - [email protected] * Corresponding author
Published: 24 August 2004 Genetic Vaccines and Therapy 2004, 2:11
doi:10.1186/1479-0556-2-11
Received: 13 May 2004 Accepted: 24 August 2004
This article is available from: http://www.gvt-journal.com/content/2/1/11 © 2004 Harms et al; licensee BioMed Central Ltd. This is an open-access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract Background: Viral promoters are used in mammalian expression vectors because they generally have strong activity in a wide variety of cells of differing tissues and species. Methods: The utility of the BLV LTR/promoter (BLVp) for use in mammalian expression vectors was investigated through direct comparison to the CMV promoter (CMVp). Promoter activity was measured using luciferase assays of cell lines from different tissues and species stably transduced with BLVp or CMVp driven luciferase vectors including D17, FLK, BL3.1 and primary bovine B cells. Cells were also modified through the addition of BLV Tax expression vectors and/or BLV infection as well as treatment with trichostatin A (TSA). Results: Results indicate the BLV promoter, while having low basal activity compared to the CMV promoter, can be induced to high-levels of activity similar to the CMV promoter in all cells tested. Tax or BLV infection specifically enhanced BLVp activity with no effect on CMVp activity. In contrast, the non-specific activator, TSA, enhanced both BLVp and CMVp activity. Conclusion: Based on these data, we conclude the BLV promoter could be very useful for transgene expression in mammalian expression vectors.
Background Viral promoters are commonly used as regulatory elements in gene therapy vectors due to their strong activity in various cell lines in vitro. Probably the most widely used promoter in mammalian expression systems is the human cytomegalovirus immediate-early gene (CMV) promoter. The CMV promoter induces high-level constitutive expression in a variety of mammalian cell lines [1].
In many gene therapy applications, however, an inducible or cell specific promoter would be more appropriate. A regulated transgene expression sy
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