Modification of Recombination-based GATEWAY<sup>TM</sup> Binary Destination Vector with Novel Promoter for <i>Agrobacterium</i>-mediated Transformation of Rice

Authors

  • Md Rakibul Islam Rice Research and Biotechnology Laboratory, Department of Biochemistry and Molecular Biology, University of Dhaka, Dhaka-1000, Bangladesh
  • Richard Malo Rice Research and Biotechnology Laboratory, Department of Biochemistry and Molecular Biology, University of Dhaka, Dhaka-1000, Bangladesh
  • Rumana S Tammi Rice Research and Biotechnology Laboratory, Department of Biochemistry and Molecular Biology, University of Dhaka, Dhaka-1000, Bangladesh
  • Sharmin Jahan Rice Research and Biotechnology Laboratory, Department of Biochemistry and Molecular Biology, University of Dhaka, Dhaka-1000, Bangladesh
  • Lisa Parvin Rice Research and Biotechnology Laboratory, Department of Biochemistry and Molecular Biology, University of Dhaka, Dhaka-1000, Bangladesh
  • Zeba I Seraj Rice Research and Biotechnology Laboratory, Department of Biochemistry and Molecular Biology, University of Dhaka, Dhaka-1000, Bangladesh

DOI:

https://doi.org/10.3329/ptcb.v17i1.1120

Keywords:

Recombination, Binary vector, Promoter, Transformation, Rice

Abstract

The GATEWAYTM Binary Destination Vector pH7WG2 is available for easy insertion of genes for transformation into plants. The gene of interest integrates downstream of the Cauliflower Mosaic Virus Promoter CaMV 35S by recombination. The CaMV 35S promoter is however not suitable for transformation and expression of genes in monocots like rice. We isolated and cloned a ~1100 bp upstream region from two rice (Pokkali and IR64) Na/H antiporter genes into the GATEWAYTM promoter cloning vector pHGWFS7. The Pokkali promoter expressed the â-glucuronidase or GUS gene ~25-fold more efficiently than the CaMV 35S promoter in rice calli, while that of IR64 was 7-fold more. The IR64 promoter however showed efficient expression in transgenic rice leaves. The promoter from Pokkali Na/H antiporter was used to replace the CaMV 35S sequence in pH7WG2. The CaMV 35S region was cut out and the linear vector fragment blunted and T-tailed. After amplification of the promoter from Pokkali rice DNA, it was A-tailed and ligated to the modified T-vector. The resultant vector, named pH7WG3, following the nomenclature at the gateway site, www.plantgenetics.rug.ac.be/gateway, can now be used for recombination of any genes for efficient rice transformation.

Key words: Recombination, Binary vector, Promoter, Transformation, Rice

DOI = 10.3329/ptcb.v17i1.1120

Plant Tissue Cult. & Biotech. 17(1): 47-58, 2007 (June)

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How to Cite

Islam, M. R., Malo, R., Tammi, R. S., Jahan, S., Parvin, L., & Seraj, Z. I. (2008). Modification of Recombination-based GATEWAY<sup>TM</sup> Binary Destination Vector with Novel Promoter for <i>Agrobacterium</i>-mediated Transformation of Rice. Plant Tissue Culture and Biotechnology, 17(1), 47–58. https://doi.org/10.3329/ptcb.v17i1.1120

Issue

Vol. 17 No. 1 (2007)

Section

Articles