Abstract
The narB gene from the cyanobacterium Synechococcus sp. PCC 7942 was cloned downstream from the LacI-regulated promoter Ptrc in the Escherichia coli vector pTrc99A, rendering plasmid pCSLM1. Addition of isopropyl-β-D-thiogalactoside to E. coli (pCSLM1) resulted in the parallel expression of a 76 kDa polypeptide and a nitrate reductase activity with properties identical to those known for nitrate reductase isolated from Synechococcus cells. As is the case for nitrate reductase from Synechococcus cells, either reduced methyl viologen or reduced ferredoxin could be used as an electron donor for the reduction of nitrate catalyzed by E. coli (pCSLM1) extracts. This data shows that narB is a cyanobacterial structural gene for nitrate reductase.
References
Andriesse AJ, Bakker H: Synechococcus sp. nrtD and narB genes for nitrate transporter and nitrate reductase. EMBL/GenBank/DDBJ accession number X74597.
Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (ed) Currents Protocols in Molecular Biology. Greene Publishing/Wiley Inter-science, New York (1995).
Candau P: Purificación y propiedades de la ferredoxinanitrato reductasa de la cianobacteria Anacystis nidulans. Ph. D. thesis, Universidad de Sevilla, Spain (1979).
Crawford NM, Arst HNJr: The molecular genetics of nitrate assimilation in fungi and plants. Annu Rev Genet 27: 115–146 (1993).
Flores E, Herrero A: Assimilatory nitrogen metabolism and its regulation, in Bryant DA (ed) The Molecular Biology of Cyanobacteria, pp. 487–517. Kluwer Academic Publishers, Dordrecht, Netherlands (1994).
Gangeswaran R, Lowe DJ, Eady RR: Purification and characterization of the assimilatory nitrate reductase of Azotobacter vinelandii. Biochem J 289: 335–342 (1993).
Guerrero MG, Vega JM, Losada M: The assimilatory nitrate-reducing system and its regulation. Annu Rev Plant Physiol 32: 169–204 (1981).
Herrero A, Flores E, Guerrero MG: Regulation of nitrate reductase levels in the cyanobacteria Anacystis nidulans, Anabaena sp. strain 7119, and Nostoc sp. strain 6719. J Bact 145: 175–180 (1981).
Kuhlemeier CJ, Logtenberg T, Stoorvogel W, Heugten HAA, Borrias WE, van Arkel GA: Cloning of nitrate reductase genes from the cyanobacterium Anacystis nidulans. J Bact 159: 36–41 (1984).
Kuhlemeier CJ, Teeuwsen WJP, Janssen MJT, van Arkel GA: Cloning of a third nitrate reductase gene from the cyanobacterium Anacystis nidulans R2 using a shuttle gene library. Gene 31: 109–116 (1984).
Lin JT, Goldman BS, Stewart V: Structures of genes nasA and nasB, encoding assimilatory nitrate and nitrite reductases in Klebsiella pneumoniae M5al. J Bact 175: 2370–2378 (1993).
Lin JT, Goldman BS, Stewart V: The nasFEDCBA operon for nitrate and nitrite assimilation in Klebsiella pneumoniae M5al. J Bact 176: 2551–2559 (1994).
Luque I, Herrero A, Flores E, Madueño F: Clustering of genes involved in nitrate assimilation in the cyanobacterium Synechococcus. Mol Gen Genet 232: 7–11 (1992).
Luque I, Flores E, Herrero A: Nitrite reductase gene from Synechococcus sp. PCC 7942: homology between cyanobacterial and higher-plant nitrite reductases. Plant Mol Biol 21: 1201–1205 (1993).
Luque I, Flores E, Herrero A: Nitrate and nitrite transport in the cyanobacterium Synechococcus sp. PCC 7942 are mediated by the same permease. Biochim Biophys Acta 1184: 296–298 (1994).
Luque I, Flores E, Herrero A: Molecular mechanism for the operation of nitrogen control in cyanobacteria. EMBO J 13: 2862–2869 (1994).
Omata T, Andriesse X, Hirano A: Identification and characterization of a gene cluster involved in nitrate transport in the cyanobacterium Synechococcus sp. PCC 7942. Mol Gen Genet 236: 193–202 (1993).
Madueño F, Borrias WE, van Arkel GA, Guerrero MG: Isolation and characterization of Anacystis nidulans R2 mutants affected in nitrate assimilation: establishment of two new mutant types. Mol Gen Genet 213: 223–228 (1988).
Manzano C, Candau P, Gómez-Moreno C, Relimpio AM, Losada M: Ferredoxin-dependent photosynthetic reduction of nitrate and nitrite by particles of Anacystis nidulans. Mol Cell Biochem 10: 161–169 (1976).
Martín-Nieto J, Flores E, Herrero A: Biphasic kinetic behavior of nitrate reductase from heterocystous, nitrogenfixing cyanobacteria. Plant Physiol 100: 157–163 (1992).
Rajagopalan KV, Johnson JL: The pterin molybdanum cofactors. J Biol Chem 267: 10199–10202 (1992).
Schleif RF, Wensink PC: Practical Methods in Molecular Biology. Springer-Verlag, New York (1981).
Siddiqui RA, Warnecke-Eberz U, Hengsberger A, Schneider B, Kostka S, Friedrich B: Structure and fimction of a periplasmic nitrate reductase in Alcaligenes eutrophus H16. J Bact 175: 5867–5876 (1993).
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Rubio, L.M., Herrero, A. & Flores, E. A cyanobacterial narB gene encodes a ferredoxin-dependent nitrate reductase. Plant Mol Biol 30, 845–850 (1996). https://doi.org/10.1007/BF00019017
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DOI: https://doi.org/10.1007/BF00019017