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Isolation, sequence and expression in Escherichia coli of the nitrite reductase gene from the filamentous, thermophilic cyanobacterium Phormidium laminosum

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Abstract

The nitrite reductase (NiR) gene (nirA) has been isolated and sequenced from the filamentous, thermophilic non-N2-fixing cyanobacterium Phormidium laminosum. Putative promoter-like and Shine-Dalgarno sequences appear at the 5′ end of the 1533 bp long nir-coding region. The deduced amino acid sequence of NiR from P. laminosum corresponds to a 56 kDa polypeptide, a size identical to the molecular mass previously determined for the pure enzyme, and shows a high identity with amino acid sequences from ferredoxin-dependent NiR. This cyanobacterial NiR gene has been efficiently expressed in Escherichia coli DH5α from the E. coli lac promoter and probably from the P. laminosum NiR promoter.

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Abbreviations

IPTG:

isopropyl-β-D-thiogalactopyranoside

NiR:

nitrite reductase

NR:

nitrate reductase

NT:

nitrate transport

SiR:

sulfite reductase

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Authors and Affiliations

  1. Departamento de Bioquímica y Biología Molecular, Facultad de Ciencias, Universidad de Córdoba, Avda. San Alberto Magno s/n, E-14071, Córdoba, Spain

    Faustino Merchán, Rafael Prieto & Emilio Fernández

  2. Departamento de Bioquímica y Biología Molecular, Facultad de Ciencias, Universidad del País Vasco, Apartado 644, E-48080, Bilbao, Spain

    Faustino Merchán, María J. Llama & Juan L. Serra

  3. Plant Science Center, Cornell University, 139 Biotechnology Building, 14853-2703, Ithaca, NY, USA

    Karen L. Kindle

Authors
  1. Faustino Merchán
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  2. Rafael Prieto
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  3. Karen L. Kindle
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  4. María J. Llama
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  5. Juan L. Serra
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  6. Emilio Fernández
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Merchán, F., Prieto, R., Kindle, K.L. et al. Isolation, sequence and expression in Escherichia coli of the nitrite reductase gene from the filamentous, thermophilic cyanobacterium Phormidium laminosum . Plant Mol Biol 27, 1037–1042 (1995). https://doi.org/10.1007/BF00037030

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  • DOI: https://doi.org/10.1007/BF00037030

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