SpyTag can spontaneously form a covalent isopeptide bond with its protein partner SpyCatcher. Firefly luciferase from Photinus pyralis was cyclized in vivo by fusing SpyCatcher at the N terminus and SpyTag at the C terminus. Circular LUC was more thermostable and alkali-tolerant than the wild type, without compromising the specific activity. Structural analysis indicated that the cyclized LUC i...

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.

Bioluminescence imaging (BLI) is emerging as a powerful tool for real-time monitoring of infections in living animals. However, since luciferases are oxygenases, it has been suggested that the requirement for oxygen may limit the use of BLI in anaerobic environments, such as the lumen of the gut. Strains of Escherichia coli harboring the genes for either the bacterial luciferase from Photorhabd...

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...

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...

We studied adenosine triphosphate (ATP) synthesized by bacteria in tube dilution cultures to devise a rapid antibacterial susceptibility test. The ATP level was determined with a firefly (Photinus pyralis) luciferin/luciferase bioluminescence assay, which allows a direct quantifying of bacteria above 10,000 CFU/ml. At sub-MIC, total bacterial ATP levels were paralleled by increased viable bacte...