Cloning and heterologous expression of nitrate reductase genes from Dunaliella salina

摘要: A cDNA corresponding to the nitrate reductase (NR) gene from Dunaliella salina was isolated by RT-PCR and (5′/3′)-RACE techniques. The full-length sequence of 3,694 bp contained an open reading frame 2,703 encoding 900 amino acids, a 5′-untranslated region 151 3′-untranslated 840 with poly (A) tail. putative product exhibited 78%, 65%, 59% 50% identity in acid genes tertiolecta, Volvox carteri, Chlamydomonas reinhardtii, Chlorella vulgaris, respectively. Phylogenetic analysis showed that D. NR clusters together known proteins green algae. molecular mass encoded protein predicted be 99.5 kDa, isoelectric point 8.31. This shares common structural features NRs higher plants heterologously expressed Escherichia coli as fusion protein, accumulated up 21% total bacteria protein. Recombinant active enzyme assay, confirming cloned is indeed NR.