African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12405

Full Length Research Paper

Cloning and expression of a tomato glutathione S- transferase (GST) in Escherichia coli

  Dongli Pei1, Jinping Ding1, Zilei Duan1, Meng Li1, Yiyang Feng1 and Chengwei Li1,2*        
  1Key laboratory of Plant-Microbe Interaction, Department of Life Science, Shangqiu Normal University, Shangqiu 476000, China. 2Key Laboratory of Plant Genetics and Molecular Breeding, Department of Life Science, Zhoukou Normal University, Zhoukou 466001, China.
Email: [email protected]

  •  Accepted: 01 March 2012
  •  Published: 20 March 2012

Abstract

 

Glutathione S- transferases (GSTs) fulfill a diverse range of functions in an organism. In a previous study, a putative glutathione S-transferase gene (ShGSTU1) from a wild-type tomato, Solanum habrochaites G1.1560, was identified to be a key gene in pathogen resistant response against powdery mildew in tomato. In this study,ShGSTU1 was cloned into plasmid pET-28a, efficiently expressed in Escherichia coliupon isopropyl-β-D-1-thiogalactopyronoside (IPTG) induction, purified with Ni2+ affinity chromatography and biochemically characterized. The results show that the optimal conditions for the expression of recombinant ShGSTU1 in E. coli were growth under 37°C, and 4-h IPTG induction with 1 mM concentration. About 18.93 mg ShGSTU1 was recovered from 1 g wet bacteria. The recombinant ShGSTU1 exhibited enzymatic activity with specific activity 0.625 U/mg. These results might provide a significant foundation for the later research on the mechanism of ShGSTU1 in tomato resistance to powdery mildew.

 

Key words: Tomato, glutathione S-transferase, expression, purification, enzyme activity.

Abbreviation

GSTs, Glutathione S- transferases; IPTG, isopropyl-β-D-1-thiogalactopyronoside.