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Cloning, molecular characterization and heterologous expression of a glutathione S-transferase gene in rice

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Abstract

OsGSTL2 is one of three tandem-arranged glutathione S-transferase, lambda class genes in chromosome 3 of rice (Oryza sativa L.). It includes 9 introns and 10 exons, and encodes a protein of 244 amino acid residues with a calculated molecular mass of 27.37 kDa. The predicted three-dimensional structure of OsGSTL2 showed a typical glutathione S-transferase fold. Using semi-quantitative RT-PCR analysis, OsGSTL2 transcript was detected in the roots and leaves of seedling stage and tillering stage, and the roots, leaves and panicles of heading stage from rice plants, and the expression level of OsGSTL2 mRNA in rice roots show significant change under chlorsulfuron stress. The OsGSTL2 gene was cloned into pYTV vector and was transformed into yeast strain PEP4. Western blot analysis showed the exogenous OsGSTL2 was expressed in transformed yeast. GST activity of crude extracts of yeast showed the OsGSTL2 transgenic yeast had higher levels of GST activities than control yeasts. These findings suggested that the OsGSTL2 is a glutathione S-transferase and has potential use in detoxification.

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Abbreviations

ABA:

abscisic acid

CDNB:

1-chloro-2,4-dinitrobenzene

2-CBSU:

N-(aminocarbonyl)-2-chlorobenzenesulfonamide

DHAR:

dehydroascorbate reductase

ET:

ethylene

GST:

glutathione S-transferases

GSTF:

phi class GST

GSTL:

lambda class GST

GSTT:

theta class GST

GSTU:

tau class GST

GSTZ:

zeta class GST

JA:

jasmonic acid

MAPEG:

membrane-associated proteins in eicosanoid and glutathione metabolism

PCR:

polymerase chain reaction

ROS:

reactive oxygen species

SA:

salicylic acid

TCHQD:

tetrachlorohydroquinone dehalogenase

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Authors and Affiliations

  1. Key Laboratory of Biorheological Science and Technology, Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, 400044, China

    T. Z. Hu, S. He, L. Deng & G. X. Wang

  2. Department of Biology, Chongqing Three Gorges University, Chongqing, 404000, China

    T. Z. Hu & X. Y. Huang

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  1. T. Z. Hu
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  2. S. He
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  3. X. Y. Huang
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  4. L. Deng
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  5. G. X. Wang
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Correspondence to T. Z. Hu.

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Hu, T.Z., He, S., Huang, X.Y. et al. Cloning, molecular characterization and heterologous expression of a glutathione S-transferase gene in rice. Russ J Bioorg Chem 37, 344–350 (2011). https://doi.org/10.1134/S1068162011030174

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