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

  • Mark Tepfer (a1), Stéphane Gaubert (a1), Mathieu Leroux-Coyau (a2), Sonia Prince (a2) and Louis-Marie Houdebine (a2)...

Abstract

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.

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References

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[1] 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
[2] Assaad FF, Signer ER (1990) Cauliflower mosaic virus P35S promoter activity in Escherichia coli. Mol. Gen. Genet. 223: 517–520
[3] 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
[4] 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
[5] 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
[6] 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
[7] 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
[8] 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
[9] Dennis E, Berg P (1985) Transcription from a plant promoter in animal cells. Nucl. Acids Res. 13: 7945–7957
[10] 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: 141150
[11] 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: 763773
[12] 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
[13] Ho MW, Ryan A, Cummins J (1999) Cauliflower mosaic viral promoter – recipe for disaster? Microb. Ecol. Health Dis. 11: 194–197
[14] 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
[15] Hochheimer A, Tjian R (2003) Diversified transcription initiation complexes expand promoter selectivity and tissue-specific gene expression. Genes Dev. 17: 1300–1320
[16] Hull R (2002) Mathews’ Plant Virology. Academic Press. San Diego, San Francisco, New York, Boston, London, Sydney, Tokyo
[17] 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
[18] 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
[19] 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
[20] 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: 18711876
[21] Langridge, P, Feix, G (1983) A zein gene of maize is transcribed from two widely separated promoter regions. Cell 34: 10151022
[22] Martinez E (2002) Multi-protein complexes in eukaryotic gene transcription. Plant Mol. Biol. 50: 925–947
[23] Morel, JB, Tepfer, M (2000) Pour une évaluation scientifique des risques : le cas du promoteur 35S. Biofutur 201: 3235 (available in English from Mark.Tepfer@versailles.inra.fr).
[24] Odell, JT, Nagy, F, Chua, NH (1985) Identification of DNA sequences required for activity of the cauliflower mosaic virus 35S promoter. Nature 313: 810812
[25] 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
[26] 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
[27] 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
[28] 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
[29] 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
[30] 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
[31] 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
[32] 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
[33] 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
[34] 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

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

  • Mark Tepfer (a1), Stéphane Gaubert (a1), Mathieu Leroux-Coyau (a2), Sonia Prince (a2) and Louis-Marie Houdebine (a2)...

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