Abstract
This study explored the possibility of using non-viral, plant-based gene sequences to create strong and constitutive expression vectors. Replacement histone H3 genes are highly and constitutively expressed in all plants. Sequences of the cloned alfalfa histone H3.2 gene MsH3g1 were tested. Constructs of the β-glucuronidase (GUS) reporter gene were produced with H3.2 gene promoter and intron sequences. Their efficiency was compared with that of the commonly used strong 35S cauliflower mosaic virus promoter in transgenic tobacco plants. Combination of the H3.2 promoter and intron produced significantly higher GUS expression than the strong viral 35S promoter. Histochemical GUS analysis revealed a constitutive pattern of expression. Thus, alfalfa replacement H3 gene sequences can be used instead of viral promoters to drive heterologous gene expression in plants, avoiding perceived risks of viral sequences.
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References
Al-Kaff NS, Covey SN, Kreike MM, Page AM, Pinder R and Dale PJ (1998) Transcriptional and posttranscriptional plant gene silencing in response to a pathogen. Science 279: 2113–2115.
Al-Kaff NS, Kreike MM, Covey SN, Pitcher R, Page AM and Dale PJ (2000) Plants rendered herbicide-susceptible by cauliflower mosaic virus-elicited suppression of a 35S promoter-regulated transgene. Nat Biotechnol 18: 995–999.
Bevan M (1984) Binary Agrobacterium vectors for plant transformation. Nucl Acids Res 12: 8711–8721.
Fisher DK and Guiltinan MJ (1995) Rapid, efficient production of homozygous transgenic tobacco plants with Agrobacterium tumefaciens: a seed-to-seed protocol. Plant Mol Biol Rep 13: 278–289.
Holsters M, de Waele D, Depicker A, Messens E, Van Montagu M and Schell J (1978) Transfection and transformation of A. tumefaciens. Mol Gen Genet 163: 181–187.
Kapros T, Stefanov I, Magyar Z, Ocsovszky I and Dudits D (1993) A short histone H3 promoter from alfalfa specifies expression in S phase cells and meristems. In Vitro Cell Dev Biol 29: 27–32.
Kapros T, Robertson AJ and Waterborg JH (1995) Histone H3 transcript stability in alfalfa. Plant Mol Biol 28: 901–914.
Kohli A, Griffiths S, Palacios N, Twyman RM, Vain P, Laurie DA and Christou P (1999) Molecular characterization of transforming plasmid rearrangements in transgenic rice reveals a recombination hotspot in the CaMV 35S promoter and confirms the predominance of microhomology mediated recombination. Plant J 17: 591–601.
Marzluff WF (1989) Histone gene expression not coupled to DNA synthesis. In: Hnilica LS, Stein GS and Stein LJ (eds.), Histones and Other Basic Nuclear Proteins. (pp. 319–329) CRC Press, Boca Raton.
Robertson, AJ, Kapros T, Dudits D and Waterborg JH (1996) Identification of three highly expressed replacement histone H3 genes of alfalfa. DNA Seq 6: 137–146.
Sambrook J, Fritsch EF and Maniatis T (1989) Molecular Cloning. A Laboratory Manual. 2nd edn. Cold Spring Harbor, NY.
Szabados L, Ratet P, Grunenberg B and de Bruijn FJ (1990) Functional analysis of the Sesbania rostrata leghemoglobin glb3 gene 5′-upstream region in transgenic Lotus corniculatus and Nicotiana tabacum plants. Plant Cell 10: 973–986.
Waterborg JH and Robertson AJ (1996) Common features of analogous replacement histone H3 genes in animals and plants. J Mol Evol 43: 194–206.
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Kelemen, Z., Mai, A., Kapros, T. et al. Transformation vector based on promoter and intron sequences of a replacement histone H3 gene. A tool for high, constitutive gene expression in plants. Transgenic Res 11, 69–72 (2002). https://doi.org/10.1023/A:1013923826979
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DOI: https://doi.org/10.1023/A:1013923826979