Isolation and characterization of the nitrate reductase structural gene of Chlamydomonas reinhardtii.

• DOI: 10.1073/pnas.86.17.6449
• Publication Date: 2006-05-31T11:02:09Z
• Authors: E Fernández; R Schnell; L P Ranum; S C Hussey; C D Silflow; P A Lefebvre
• Keywords: Chlamydomonas reinhardtii; Structural gene

The nitrate reductase structural gene of Chlamydomonas reinhardtii has been isolated from a genomic library by using cDNA probe barley. Restriction fragment length polymorphism analyses mapped the clone (B6a) to locus nit-1. Overlapping inserts cover region genome about 24 kilobases containing entire gene, which spans approximately 5-8 kilobases. Sequence analysis DNA fragments B6a demonstrated high degree sequence similarity at amino acid level with regions corresponding portions heme and FAD/NADH-binding domains tobacco Arabidopsis thaliana reductases human NADH cytochrome b5 reductase. identity cloned as was confirmed its ability complement nit-1 mutation upon transformation. produced 3.4-kilobase transcript in cells derepressed nitrate; undetectable grown presence ammonium. In that contain putative regulatory nit-2, significantly lower levels were found, indicating wild-type nit-2 is involved control levels.