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KMID : 0380220060390050578
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Journal of Biochemistry and Molecular Biology
2006 Volume.39 No. 5 p.578 ~ p.585
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cDNA Cloning, Expression and Homology Modeling of a Luciferase from the Firefly Lampyroidea maculata
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Emamzadeh Abdo Rahman
Hosseinkhani Saman
Sadeghizadeh Majid
Nikkhah Maryam
Chaichi Mohammad Javad
Mortazavi Mojtaba
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Abstract
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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 AMPbinding 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.
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KEYWORD
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AMP-Binding domain, Firefly, Homology modeling, Lampyroidea maculata, Luciferase, Sequence analysis
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