African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12476

Full Length Research Paper

Overexpression of an abiotic-stress inducible plant protein in the bacteria Escherichia coli

Aryadeep Roychoudhury1, 2* and Supratim Basu1
  1Department of Botany, Bose Institute, 93/1, Acharya Prafulla Chandra Road, Kolkata-700009, West Bengal, India. 2Department of Botany, Plant Molecular Biology and Biotechnology Laboratory, University of Calcutta, 35, Ballygunge Circular Road, Kolkata- 700 019, West Bengal, India.
Email: [email protected]

  •  Accepted: 15 August 2008
  •  Published: 17 September 2008

Abstract

 

The aim of our work was the overexpression of the abiotic stress-inducible dehydrin protein, namely RAB16A, from rice in the BL21 strain of Escherichia coli. The Rab16Atranscript of 0.5 Kbp was amplified from the total RNA of the salt-tolerant indica rice cultivar Nonabokra by RT-PCR and cloned into the expression vector pGEX-3X. The 47 kDa protein, expressed as GST: RAB16A fusion protein, after 2 mM IPTG-mediated induction, was collected as S10 fraction and purified through glutathione-sepharose affinity resin. Immunoblot analysis with the maize dehydrin antiserum showed cross-reaction with the above band, but not with GST protein alone, showing functional expression of the heterologous RAB16A protein in the bacterial system.

 

Key words:  Fusion protein, Glutathione-sepharose, GST: RAB16A, pGEX-3X.