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

  • A Roychoudhury
  • S Basu
Keywords: Fusion protein, Glutathione-sepharose, GST, RAB16A, pGEX-3X.

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 Rab16A transcript 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 glutathionesepharose affinity resin. Immunoblot analysis with the maize dehydrin antiserum showed crossreaction with the above band, but not with GST protein alone, showing functional expression of the heterologous RAB16A protein in the bacterial system.
Section
Articles

Journal Identifiers


eISSN: 1684-5315