5′ Regulatory region of ubiquitin 2 gene from Porteresia coarctata makes efficient promoters for transgene expression in monocots and dicots
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Porteresia ubiquitin 5′ regulatory region drives transgene expression in monocots and dicots.
Ubiquitin promoters are promising candidates for constitutive transgene expression in plants. In this study, we isolated and characterized a novel 5′ regulatory sequence of a ubiquitin gene from Porteresia coarctata, a stress-tolerant wild grass species. Through functional analysis in heterologous plant systems, we have demonstrated that full length (Port Ubi2.3) or truncated sequence (PD2) of the isolated regulatory fragment can drive constitutive expression of GUS in monocots and/or dicots. In silico analysis of Port Ubi2.3 has revealed the presence of a 640 bp core promoter region followed by two exons and two introns with numerous putative cis-acting sites scattered throughout the regulatory region. Transformation and expression studies of six different deletion constructs in rice, tobacco and sugarcane revealed that the proximal intron has an enhancing effect on the activity of the core promoter in both monocots and dicots, whereas, Port Ubi2.3 was able to render strong expression only in monocots. This regulatory sequence is quite distinct from the other reported ubiquitin promoters in structure and performs better in monocots compared to other commonly used promoters—maize Ubi1 and Cauliflower Mosaic Virus 35S.
KeywordsUbiquitin promoter Porteresia coarctata Ubi 5′ regulatory region Transgenic monocot Transgenic dicot Promoter for plant transformation
The authors would like to thank Indian Council of Agricultural Research and Sugarcane Breeding Institute, Coimbatore for the funding and infrastructure.
- Arvinth S, Arun S, Selvakesavan RK, Srikanth J, Mukunthan N, Ananda Kumar P, Premachandran MN, Subramonian N (2010) Genetic transformation and pyramiding of aprotinin-expressing sugarcane with cry1Ab for shoot borer (Chilo infuscatellus) resistance. Plant Cell Rep 29:383–395PubMedCrossRefGoogle Scholar
- Buchanan BB, Gruissem W, Jones RL (2000) Biochemistry and molecular biology of plants. American Society of Plant Physiologists, Rockville, pp 340–342Google Scholar
- Doyle JJ, Doyle JL (1990) Isolation of plant DNA from fresh tissue. Focus 12:13–15Google Scholar
- Gowik U, Burscheidt J, Akyildiz M, Schlue U, Koczor M, Streubel M, Westhoff P (2004) Cis-regulatory elements for mesophyll-specific gene expression in the C4 plant Flaveria trinervia, the promoter of the C4 phosphoenolpyruvate carboxylase gene. Plant Cell 23:2087–2105Google Scholar
- Horsch RB, Fry J, Hoffmann N, Neidermeyer J, Rogers SG, Fraley RT (1988) Leaf disc transformation. In: Gelvin SB, Schilperoort RA (eds) Plant molecular biology manual. Kluwer Academic Publishers, Dordrecht, pp 1–9Google Scholar
- Kuriakose B, Ganesan V, Thomas G, Viswanathan A, Anand N (2009b) Random amplification of genomic ends (RAGE) as an efficient method for isolation and cloning of promoters and uncloned genomic regions. Afr J Biotechnol 8:4765–4773Google Scholar
- Mitsuhara I, Ugaki M, Hirochika H, Ohshima M, Murakami T, Gotoh Y, Katayose Y, Nakamura S, Honkura R, Nishimiya S, Ueno K, Mochizuki A, Tanimoto H, Tsugawa H, Otsuki Y, Ohashi Y (1996) Efficient promoter cassettes for enhanced expression of foreign genes in dicotyledonous and monocotyledonous plants. Plant Cell Physiol 37:49–59PubMedCrossRefGoogle Scholar
- Potenza C, Aleman L, Sengupta-Gopalan C (2004) Targeting transgene expression in research, agricultural and environmental applications: promoters used in plant transformations. In Vitro Cell Dev Biol Plant 40:1–22Google Scholar