NDLI: Cloning and heterologous expression of nitrate reductase genes from Dunaliella salina

Content Provider	Springer Nature Link
Author	Xie, Hua Xu, Peirong Jia, Yanlong Li, Jie Lu, Yumin Xue, Lexun
Copyright Year	2007
Abstract	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 cDNA sequence of 3,694 bp contained an open reading frame of 2,703 bp encoding 900 amino acids, a 5′-untranslated region of 151 bp and a 3′-untranslated sequence of 840 bp with a poly (A) tail. The putative gene product exhibited 78%, 65%, 59% and 50% identity in amino acid sequence to the corresponding genes of Dunaliella tertiolecta, Volvox carteri, Chlamydomonas reinhardtii, and Chlorella vulgaris, respectively. Phylogenetic analysis showed that D. salina NR clusters together with known NR proteins of the green algae. The molecular mass of the encoded protein was predicted to be 99.5 kDa, with an isoelectric point of 8.31. This protein shares common structural features with NRs from higher plants and green algae. The full-length cDNA was heterologously expressed in Escherichia coli as a fusion protein, and accumulated to up to 21% of total bacteria protein. Recombinant NR protein was active in an enzyme assay, confirming that the cloned gene from D. salina is indeed NR.
Starting Page	497
Ending Page	504
Page Count	8
File Format	PDF
ISSN	09218971
Journal	Journal of Applied Phycology
Volume Number	19
Issue Number	5
e-ISSN	15735176
Language	English
Publisher	Springer Netherlands
Publisher Date	2007-03-01
Publisher Place	Dordrecht
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Cloning Dunaliella salina Chlorophyta Nitrate reductase Overexpression Ecology Hydrobiology Plant Sciences Plant Physiology
Content Type	Text
Resource Type	Article
Subject	Plant Science Aquatic Science

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