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Molecular cloning and characterisation of cytoplasmic glutamine synthetase gene BcGS1 from non‐heading Chinese cabbage

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BACKGROUND: Glutamine synthetase (GS; EC 6.3.1.2) is a key enzyme of nitrogen (N) assimilation, catalysing the synthesis of glutamine from ammonium and glutamate. Plants have two types of GS isoenzyme that are localised in different compartments: one in the cytosol (GS1) and the other in the chloroplast (GS2). GS1 is the major form of GS in plant roots and directly converts ammonium taken up by plant roots to glutamine.

RESULTS: The GS1 gene cDNA of non‐heading Chinese cabbage (Brassica campestrisssp. chinensis Makino) cultivar ‘Suzhouqing’ was isolated by RT‐PCR (real‐time polymerase chain reaction) and (5′/3′)‐RACE (rapid amplification of cDNA ends) techniques. It was classified as GS1 by sequence alignment and motif search and named B. campestris ssp. chinensis Makino GS1 (BcGS1). Subcellular localisation analysis showed that BcGS1 was distributed in the cytoplasm of cells. BcGS1 was expressed in all parts, but mainly in the roots, which was verified by northern blotting analysis. Additionally, its expression was influenced by the N source concentration.

CONCLUSION: These results suggest that BcGS1 is a novel member of the GS family in plants. BcGS1 was significantly related to N assimilation in non‐heading Chinese cabbage, demonstrating that this gene plays an important role in plant growth and development. Copyright © 2010 Society of Chemical Industry
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Keywords: Brassica campestris ssp. chinensis Makino; cloning; cytoplasmic glutamine synthetase; expression; subcellular localisation

Document Type: Research Article

Publication date: April 15, 2010

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