Characterisation of pea cytosolic glutathione reductase expressed in transgenic tobacco |
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Authors: | Rebecca G Stevens Gary P Creissen Philip M Mullineaux |
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Institution: | (1) John Innes Centre, Colney, Norwich, NR4 7UH, UK, GB |
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Abstract: | Expression in transgenic tobacco (Nicotiana tabacum L.) of a pea (Pisum sativum L.) GOR2 cDNA, encoding an isoform of glutathione reductase (GOR2), resulted in a 3- to 7-fold elevation of total foliar glutathione
reductase (GR) activity. The enzyme encoded by GOR2 was confirmed to be extraplastidial in organelle fractionation studies and was considered most likely to be localised in
the cytosol. A partial purification of GOR2 was achieved but a standard affinity chromatography step, using adenosine-2′,5′-diphosphate-Sepharose
and often employed in the purification of GR from diverse sources, was unsuccessful with this isoform. Preparative isoelectric
focussing was employed as part of the purification procedure of GOR2 and a complete separation from plastidial/mitochondrial
glutathione reductase (GOR1) was achieved. The isoform GOR2 was shown to have a slower migration on non-denaturing polyacrylamide
gels compared with GOR1 and properties typical of GR enzymes from plant sources.
Received: 9 November 1999 / Accepted: 28 February 2000 |
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Keywords: | : Glutathione reductase – Enzyme characterisation (leaf GR) – Nicotiana (glutathione reductase) – Pisum (GOR2 cDNA) – Transgenic tobacco |
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