Cloning and Molecular Characterization of the cDNA for the Brazilian Larval Click-beetle Pyrearinus termitilluminans Luciferase

Autor: Gonzalo L. Pérez, Fernando C. Reinach, Etelvino J. H. Bechara, Ana C. R. da Silva, Roberto Vicente Santelli, Vadim R. Viviani
Rok vydání: 1999
Předmět:
Zdroj: Photochemistry and Photobiology. 70:254-260
ISSN: 1751-1097
0031-8655
DOI: 10.1111/j.1751-1097.1999.tb07997.x
Popis: The larval click-beetle Pyrearinus termitilluminans elicits the phenomenon of luminous termite mounds in the central-west region of Brazil. The bioluminescence (BL) spectrum of this larva (Λmax= 534 run) is one of the most blue-shifted reported among known luminescent Cole-optera. We have isolated mRNA from larval thoracic lanterns and constructed a cDNA library into a δZAP II vector. An expression library was obtained after excision of the pBluescript plasmid. This library was screened by photodetection and one clone that emitted green BL (Δmax= 538 nm) was isolated. The 2.2 kb cDNA insert includes a 543 residue open reading frame showing 82% homology with the luciferase isoenzymes of Pyrophorus pla-giophthalamus (Coleoptera: Elateridae). As expected, the region between residues 223 and 247 that contains the putative active site for BL color determination showed a higher degree of homology among click-beetle luciferases that elicit closer BL colors. The in vitro BL spectrum of recombinant P. termitilluminans luciferase also peaks at 538 nm and, as in the case of native enzyme, does not show any bathochromic shift upon decreasing the pH.
Databáze: OpenAIRE
Externí odkaz: