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Cloning, functional expression and expression studies of the nitrate transporter gene from Chlorella sorokiniana (strain 211-8k)

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Abstract

The nitrate transporter from Chlorella sorokiniana (accession number AY026523) has been cloned by screening a cDNA library based on mRNA isolated after 30 min treatment of Chlorella with 5 mM nitrate and with a RT-PCR product (730 bp) as a probe. The Chlorella sequence has similarity to known nitrate transporters of the NRT2 family (high-affinity nitrate transporters). The cDNA clone was used for functional expression in Xenopus oocytes and a nitrate-dependent current was measured at pH 5.5 but not at pH 7.4. A second algal gene or a second gene product was not needed for functional expression in Xenopus. Inhibitor studies in Chlorella indicated that protein phosphorylation/dephosphorylation is involved in nitrate induction of ChNRT2.1. In addition to nitrate, ChNRT2.1 expression is induced by nitroprusside, a NO donor, and is affected by glucose.

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Authors and Affiliations

  1. Albrecht von Haller Institut für Pflanzenwissenschaften, University of Göttingen, Untere Karspuele 2, 37073, Göttingen, Germany

    Michael Koltermann, Daniela Nowara & Rudolf Tischner

  2. Instituto di Biofisica CNR-Dipartimento di Biologia, Università degli Studi di Milano, Via Celoria 26, 20133, Milano, Italy

    Anna Moroni & Sabrina Gazzarini

Authors
  1. Michael Koltermann
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  2. Anna Moroni
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  3. Sabrina Gazzarini
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  4. Daniela Nowara
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  5. Rudolf Tischner
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Correspondence to Rudolf Tischner.

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Koltermann, M., Moroni, A., Gazzarini, S. et al. Cloning, functional expression and expression studies of the nitrate transporter gene from Chlorella sorokiniana (strain 211-8k). Plant Mol Biol 52, 855–864 (2003). https://doi.org/10.1023/A:1025024821832

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