Hostname: page-component-586b7cd67f-r5fsc Total loading time: 0 Render date: 2024-11-23T03:22:56.605Z Has data issue: false hasContentIssue false
  • English
  • Français

Transient expression in mammalian cells of transgenes transcribed from the Cauliflower mosaic virus 35S promoter

Published online by Cambridge University Press:  15 September 2004

Mark Tepfer ,
Stéphane Gaubert ,
Mathieu Leroux-Coyau ,
Sonia Prince  and
Louis-Marie Houdebine
Mark Tepfer
Affiliation:
Laboratoire de Biologie Cellulaire, INRA-Versailles, 78026 Versailles Cedex, France
Stéphane Gaubert
Affiliation:
Laboratoire de Biologie Cellulaire, INRA-Versailles, 78026 Versailles Cedex, France
Mathieu Leroux-Coyau
Affiliation:
Unité de Biologie du Développement et Reproduction, INRA, 78352 Jouy-en-Josas Cedex, France
Sonia Prince
Affiliation:
Unité de Biologie du Développement et Reproduction, INRA, 78352 Jouy-en-Josas Cedex, France
Louis-Marie Houdebine
Affiliation:
Unité de Biologie du Développement et Reproduction, INRA, 78352 Jouy-en-Josas Cedex, France

Abstract

Core share and HTML view are not available for this content. However, as you have access to this content, a full PDF is available via the ‘Save PDF’ action button.

Gene constructs containing the Cauliflower mosaic virus (CaMV) 35S promoter and a sequence coding either for a green fluorescent protein (GFP) or for firefly luciferase were transfected into Chinese hamster ovary (CHO) cells. Both reporter genes were expressed to significant levels. The 35S promoter was 40 times less active than the human eF1α promoter, which is known to be one of the most potent promoters in mammalian cells. The 35S promoter must therefore be considered to be a promoter of significant potency in mammalian cells. RT-PCR analysis suggested that transcription initiation in CHO cells occurred between the TATA box and the transcription start site of the 35S promoter that function in plant cells. Further analysis by 5’RACE confirmed that transcription was initiated in CHO cells at different sites located essentially between the TATA box and the plant transcription start site, showing that 35S promoter activity in animal cells is due to the presence of promoter elements that are functional in mammalian cells, but that are not those used in plants. The data reported here raise the possibility that genes controlled by the 35S promoter, which is commonly used in transgenic plants, have the potential for expression in animal cells.

Keywords

CaMV35S promotermammalian cellstranscription
Type
Research Article
Information
Environmental Biosafety Research , Volume 3 , Issue 2 , April 2004 , pp. 91 - 97
Copyright
© ISBR, EDP Sciences, 2004

References

An G (1986) Development of plant promoter expression vectors and their use for analysis of differential activity of nopaline synthase promoter in transformed tobacco cells. Plant Physiol. 81: 86–91
Assaad FF, Signer ER (1990) Cauliflower mosaic virus P35S promoter activity in Escherichia coli. Mol. Gen. Genet. 223: 517–520
Ballas N, Broido S, Soreq H, Loyter A (1989) Efficient functioning of plant promoters and poly(A) sites in Xenopus oocytes. Nucl. Acids Res. 17: 7891–7903 CrossRef
Benfey PN, Ren L, Chua NH (1989) The CaMV 35S enhancer contains at least two domains which can confer different developmental and tissue-specific expression patterns. EMBO J. 8: 2195–2202
Burke C, Yu XB, Marchitelli L, Davis EA, Ackerman S (1990) Transcription factor IIA of wheat and human function similarly with plant and animal viral promoters. Nucl. Acids Res. 18: 3611–3620 CrossRef
Butler JEF, Kadonaga JT (2002) The RNA polymerase II core promoter: a key component in the regulation of gene expression. Genes Dev. 16: 2583–2592
Chiu WL, Niwa Y, Zeng W, Hirano T, Kobayashi H, Sheen J (1996) Engineered GFP as a vital reporter in plants. Curr. Biol. 6: 325–330 CrossRef
Cooke R, Penon P (1990) In vitro transcription from cauliflower mosaic virus promoters by a cell-free extract from tobacco cells. Plant Mol. Biol. 14: 391–405
Dennis E, Berg P (1985) Transcription from a plant promoter in animal cells. Nucl. Acids Res. 13: 7945–7957 CrossRef
Fang, RX, Nagy, F, Sivasubramaniam, S, Chua, NH (1989) Multiple cis regulatory elements for maximal expression of the cauliflower mosaic virus 35S promoter in transgenic plants. Plant Cell 1: 141–150 CrossRef
Guilley, H, Dudley, RK, Jonard, J, Balázs, E, Richards, KE (1982) Transcription of cauliflower mosaic virus DNA: detection of promoter sequences, and characterization of transcripts. Cell 30: 763–773 CrossRef
Hirt H, Kogl M, Murbacher T, Heberle-Bors E (1990) Evolutionary conservation of transcriptional machinery between yeast and plants as shown by the efficient expression from the CaMV 35S promoter and 35S terminator. Curr. Genet. 17: 473–479
Ho MW, Ryan A, Cummins J (1999) Cauliflower mosaic viral promoter – recipe for disaster? Microb. Ecol. Health Dis. 11: 194–197
Ho MW, Ryan A, Cummins J (2000) Hazards of transgenic plants containing the cauliflower mosaic virus viral promoter. Microb. Ecol. Health Dis. 12: 6–11
Hochheimer A, Tjian R (2003) Diversified transcription initiation complexes expand promoter selectivity and tissue-specific gene expression. Genes Dev. 17: 1300–1320
Hull R (2002) Mathews’ Plant Virology. Academic Press. San Diego, San Francisco, New York, Boston, London, Sydney, Tokyo
Hull R, Covey SN, Dale P (2000) Genetically modified plants and the 35S promoter; assessing the risks and enhancing the debate. Microb. Ecol. Health Dis. 12: 1–5
Katagiri F, Yamazaki KI, Horikoshi M, Roeder RG, Chua NH (1990) A plant DNA-binding protein increases the number of active preinitiation complexes in a human in vitro transcription system. Genes Dev. 4: 1899–1909
Koncz C, Kreuzaler F, Kalman ZS, Schell J (1984) A simple method to transfer, integrate and study expression of foreign genes, such as a chicken albumin and alpha-actin in plant tumors. EMBO J. 3: 1029–1037
Kunik, T, Tzfira, T, Kapulnik, Y, Gafni, Y, Dingwall, C, Citovsky, V (2001) Genetic transformation of HeLa cells by Agrobacterium. Proc. Natl. Acad. Sci. USA 98: 1871–1876 CrossRef
Langridge, P, Feix, G (1983) A zein gene of maize is transcribed from two widely separated promoter regions. Cell 34: 1015–1022 CrossRef
Martinez E (2002) Multi-protein complexes in eukaryotic gene transcription. Plant Mol. Biol. 50: 925–947
Morel, JB, Tepfer, M (2000) Pour une évaluation scientifique des risques : le cas du promoteur 35S. Biofutur 201: 32–35 (available in English from [email protected]). CrossRef
Odell, JT, Nagy, F, Chua, NH (1985) Identification of DNA sequences required for activity of the cauliflower mosaic virus 35S promoter. Nature 313: 810–812 CrossRef
Pantano T, Jolivet G, Prince S, Menck-Le Bourhis C, Maeder C, Viglietta C, Rival S, Houdebine LM (2002) Effect of the rabbit aS1-casein gene distal enhancer on the expression of a reporter gene in vitro and in vivo. Biochem. Biophys. Res. Comm. 290: 51–61
Pobjecky N, Rosenberg GH, Dinter-Gottlieb G, Kaufer D (1990) Expression of the beta-glucuronidase gene under the control of the CaMV 35S promoter in Schizosaccharomyces pombe. Mol. Gen. Genet. 220: 314–316
Prat S, Willmitzer L, Sanchez-Serrano JJ (1989) Nuclear proteins binding to a cauliflower mosaic virus 35S truncated promoter. Mol. Gen. Genet. 217: 209–214
Rüth J, Hirt H, Schweyen RJ (1992) The cauliflower mosaic virus 35S promoter is regulated by cAMP in Saccharomyces cerevisiae. Mol. Gen. Genet. 235: 365–372 CrossRef
Rüth J, Schweyen RJ, Hirt H (1994) The plant transcription factor TGA1 stimulates expression of the CaMV 35S promoter in Saccharomyces cerevisiae. Plant Mol. Biol. 25: 323–328 CrossRef
Spena A, Hain R, Ziervogel U, Saedler H, Schell J (1985) Construction of a heat-inducible gene for plants. Demonstration of heat-inducible activity of the Drosophila hsp70 promoter in plants. EMBO J. 4: 2739–2743
Sun L, Cai H, Xu W, Hu Y, Lin Z (2002) CaMV 35S promoter directs beta-glucuronidase expression in Ganoderma lucidum and Pleurotus citrinopileatus. Mol. Biotechnol. 20: 239–244 CrossRef
Taboit-Dameron F, Malassagne B, Viglietta C, Puissant C, Leroux-Coyau M, Chéreau C, Attal J, Weill B, Houdebine LM (1999) Association of the 5’HS4 sequence of the chicken $\beta$ -globin-locus control region with human EF1 $\alpha$ gene promoter induces ubiquitous and high expression of human CD55 and CD59 cDNAs in transgenic rabbits. Transgenic Res. 8: 223–235
Vlasák J, Smahel M, Pavlík A, Pavingerová D, Bríza J (2003) Comparison of hCMV immediate early and CaMV 35S promoters in both plant and animal cells. J. Biotechnol. 103: 197–202
Zahm P, Rhim SL, Geider K (1989) Promoter activity and expression of sequences from Ti-plasmid stably maintained in mammalian cells. Mol. Cell. Biochem. 90: 9–18