A cDNA library was constructed from firefly (Photinus pyralis) lantern poly(A)+ RNA, using the Escherichia coli expression vector lambda gt11. The library was screened with anti-P. pyralis luciferase (Photinus luciferin:oxygen 4-oxidoreductase, EC 1.13.12.7) antibody, and several cDNA clones expressing luciferase antigens were isolated. One clone, lambda Luc1, contained 1.5 kilobase pairs of cD...

To aid in the interpretation of high-throughput screening (HTS) results derived from luciferase-based assays, we used quantitative HTS, an approach that defines the concentration-response behavior of each library sample, to profile the ATP-dependent luciferase from Photinus pyralis against more than 70,000 samples. We found that approximately 3% of the library was active, containing only compou...

effects of atp and mg2+ concentrations on bioluminescence reaction of luciferase (photinus pyralis) were investigated by home-made luminometer. the michaelis constants of the enzyme for atp and mg2+ obtained from the lineweaver-burk graph, were 61.9 mm ±3.3 mm and 251.6mm ± 39.0mm respectively. optimum concentration of mg2+ for maximum luminescence was 0.004 m.

The Rockefeller University Press $30.00 J. Cell Biol. Vol. 185 No. 5 859–874 www.jcb.org/cgi/doi/10.1083/jcb.200812167 JCB 859 Correspondence to Anna Santamaria: [email protected] A. Uldschmid’s present address is Viramed Biotech AG, Behingerstrasse 11, D-82152, Planegg, Germany. Abbreviations used in this paper: DHC, dynein heavy chain; DIC, dynein intermediate chain; GL2, Photinus pyr...

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 phylog...

Functional expression and spectroscopic analysis of luciferases from Lampyris turkestanicus and Photinus pyralis were carried out. cDNA encoding L. turkestanicus luciferase was isolated by reverse transcription-polymerase chain reaction, cloned, and functionally expressed in Escherichia coli. The luciferases were purified to homogeneity using Ni-nitrilotriacetic acid Sepharose, and kinetic prop...

Although several enzymes known to reside in peroxisomes have been studied extensively, no cis-acting amino acid sequences involved in the transport of these proteins to peroxisomes have been described. As a first step towards the determination of a putative peroxisomal targeting sequence, we have expressed the cDNA encoding the firefly luciferase [Photinus-luciferin:oxygen 4-oxidoreductase (dec...

A rapid method for quantifying bacteriuria was evaluated in 114 clinical specimens using a bacterial adenosine triphosphate (ATP) assay. The procedure allows for removal and destruction of non-bacterial ATP and subsequent analysis of bacteria ATP by firefly (Photinus pyralis) luciferin -luciferase bioluminescence and requires 30 minutes as a whole. The correlation coefficient between the number...