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Volume 31, Issue 10
  • ISSN: 0929-8665
  • E-ISSN: 1875-5305

Abstract

Background

Bioluminescence, or the production of light by luciferases, is the basis of a well-known reporter technology. A quick way to study the efficacy of antimicrobial drugs and vaccines is bioluminescence imaging (BLI). spp. represent the only terrestrial group of bioluminescent bacteria. The luciferase obtained from has been widely used in BLI studies. However, little information is available about the functions of luciferases obtained from other members of this genera.

Objective

This study aimed to evaluate the applicability of the luciferase obtained from for BLI studies.

Methods

starE, an Iranian isolate of , was cultivated on NBTA agar plates. The resulting colonies were cultured on McConkey agar to determine the bacterial phase. Bioluminescence emission was measured using a multimode reader. The luciferase genes of this bacterium were sequenced following the PCR amplification, and the corresponding amino acid sequences were determined. The luciferase tertiary structure was then obtained from the TACOS web server and compared to that of in CE software. The operon encoding the luciferase ( genes) and substrate synthesis complex was cloned and expressed in BL21 (DE3) using the pBBR1MCS2_START vector. The luminescence emission during the growth was examined. Moreover, the effects of pH and sodium deoxycholate (bile salt) on bioluminescence emission were investigated. Appropriate conditions for the use of bioluminescent for BLI studies in mice were demonstrated in terms of cell numbers and injection routes.

Results

The bacterium was luminescent and in phase I. Its luciferase monomers (α and β) shared 100% amino acid homology with M-HU2 and more than 92% with . Tertiary structures of the luciferase monomers were 93%- 95% identical to those of . The operon was expressed in , and the maximum bioluminescence signal was observed during the decelerating phase of growth. The bioluminescence at different pH values correlated with the cell survival. The luminescence was emitted by cells exposed to the bile salt. A strong bioluminescent signal was emitted from mice after subcutaneous injection of bioluminescent at 107 CFU. However, no signals were emitted from mice that were administered the same cell number intraperitoneal injection. A 2.5-fold increase in the cell number resulted in bioluminescence detection in the abdomen of mice after intraperitoneal injection and a 3.22-fold increase in signal intensity after subcutaneous injection.

Conclusion

These results demonstrated the usefulness of luciferase for BLI studies.

© 2024 Bentham Science Publishers
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2024-10-23
2025-01-31

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/content/journals/ppl/10.2174/0109298665277974240926064439
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Characterization of Luciferase from Photorhabdus kayaii and its Application for In vivo Imaging Studies in Mice
PPL 31, 806 (2024); https://doi.org/10.2174/0109298665277974240926064439
/content/journals/ppl/10.2174/0109298665277974240926064439
/content/journals/ppl/10.2174/0109298665277974240926064439
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  • Article Type:     Research Article
Keyword(s): bile salt; Bioluminescence; luciferase; luciferase tertiary structure; Photorhabdus kayaii; TACOS

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