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Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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cDNA Cloning, Expression and Homology Modeling of a Luciferase from the Firefly Lampyroidea maculata

  • Emamzadeh, Abdo Rahman (Department of Biochemistry, Faculty of Basic Sciences, Tarbiat Modarres University) ;
  • Hosseinkhani, Saman (Department of Biochemistry, Faculty of Basic Sciences, Tarbiat Modarres University) ;
  • Sadeghizadeh, Majid (Department of Genetics, Faculty of Basic Sciences, Tarbiat Modarres University) ;
  • Nikkhah, Maryam (Institute of Biochemistry and Biophysics, University of Tehran) ;
  • Chaichi, Mohammad Javad (Department of Chemistry, Mazandaran University) ;
  • Mortazavi, Mojtaba (Department of Biochemistry, Faculty of Basic Sciences, Tarbiat Modarres University)
  • Received : 2006.04.07
  • Accepted : 2006.05.23
  • Published : 2006.09.30
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Abstract

The cDNA of a firefly luciferase from lantern mRNA of Lampyroidea maculata has been cloned, sequenced and functionally expressed. The cDNA has an open reading frame of 1647 bp and codes for a 548-residue-long polypeptide. Noteworthy, sequence comparison as well as homology modeling showed the highest degree of similarity with H. unmunsana and L. mingrelica luciferases, suggesting a close phylogenetic relationship despite the geographical distance separation. The deduced amino acid sequence of the luciferase gene of firefly L. maculata showed 93% identity to H. unmunsana. Superposition of the three-dimensional model of L. maculata luciferase (generated by homology modeling) and three dimensional structure of Photinus pyralis luciferase revealed that the spatial arrangements of Luciferin and ATP-binding residues are very similar. Putative signature of AMP-binding domain among the various firefly species and Lampyroidea maculata was compared and a striking similarity was found. Different motifs and sites have been identified in Lampyroidea maculata by sequence analysis. Expression and purification of luciferase from Lampyroidea maculata was carried out using Ni-NTA Sepharose. Bioluminescence emission spectrum was similar to Photinus pyralis luciferase.

Keywords

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