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Pristinamycin-inducible gene regulation in mycobacteria

Paper ID Volume ID Publish Year Pages File Format Full-Text
24255 43507 2009 8 PDF Available
Title
Pristinamycin-inducible gene regulation in mycobacteria
Abstract

In this work the Pip-inducible system, already used in eukaryotes, was tested in mycobacteria. This system is based on the Streptomyces coelicolor Pip repressor, the Streptomyces pristinaespiralis ptr promoter and the inducer pristinamycin I. By cloning in an integrative plasmid the ptr promoter upstream of the lacZ reporter gene and the pip gene under the control of a constitutive mycobacterial promoter, we demonstrated that the ptr promoter activity increased up to 50-fold in Mycobacterium smegmatis and up to 400-fold in Mycobacterium tuberculosis, in dependence on pristinamycin I concentration, and that the promoter was fully repressed in the absence of the inducer.Three mycobacterial genes were cloned under pptr–Pip control, both in sense and antisense direction; both proteins and antisense RNAs could be over-expressed, the antisenses causing a partial reduction of the amount of the targeted proteins. This system was used to obtain two M. tuberculosis conditional mutants in the fadD32 and pknB genes: the mutant strains grew only in the presence of the inducer pristinamycin I. Thus it showed to be an effective inducible system in mycobacteria.

Keywords
Inducible promoter; Mycobacteria; Pristinamycin; Conditional mutants
First Page Preview
Pristinamycin-inducible gene regulation in mycobacteria
Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 140, Issues 3–4, 25 March 2009, Pages 270–277
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering