12th International Symposium on Bioluminescence & Chemiluminescence

Symposium abstracts:

Generation and characterization of a bioluminescent Streptococcus pyogenes

Lamberton, G.R.¹, Pereau, M. ¹, Illes, K. ², Kelly, I.L. ¹, Childers, B.J. ³, Oberg, K.C, ^1,3, Szalay, A.A.²

(1) Department of Pathology & Human Anatomy
(2) Department of Biochemistry
(3) Department of Surgery
Loma Linda University, Loma Linda, CA. 92350, USA. Email: koberg@som.llu.edu

Introduction: S. pyogenes is a common Gram-positive organism found in the nasopharynx that can cause severe systemic and invasive disease. The development of a bioluminescent strain of S. pyogenes would aid in the study of disease progression.
Methods: Two strains of S. pyogenes were transformed with a Gram-positive plasmid, pDC123, containing a modified luciferase fusion gene, LuxF in front of the constitutive tetracycline promoter (PtetM). A Gram-positive ribosomal binding site was attached to the five prime end of the LuxF gene to allow for efficient translation in S. pyogenes. This new construct (pDC123/LuxF) was then electroporated into competent bacteria and transformants were grown on blood agar plates. Bioluminescence was characterized following exposure to the substrate decyl-aldehyde on culture plates and following subcutaneous injections into mice.
Results: Electroporation was successful in transforming both strains of S. pyogenes. Bioluminescence was first detected with 10⁵ colonies of S. pyogenes and then linearly correlated between 10⁶ and 10⁸. Detection of subcutaneous bioluminescence was about 10 fold less than on culture plates.
Conclusions: These findings demonstrate that it is possible to transform strains of S. pyogenes. Furthermore, generation of a bioluminescent strain of S. pyogenes may allow for precise monitoring of bacterial colonization, infection and disease progression in vivo.

This is a preprint of an article accepted for publication in Luminescence: Copyright 2001 John
Wiley & Sons, Ltd (Wiley website)