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Published on: December 5, 2016
Cloning, functional expression and expression studies of the nitrate transporter gene from Chlorella sorokiniana
Michael Koltermann1, Anna Moroni, Sabrina Gazzarini
1Albrecht von Haller Institut für Pflanzenwissenschaften, University of Göttingen, Untere Karspuele 2, 37073 Göttingen, Germany.
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.
