A tetracycline-inducible integrative expression system for Streptococcus pneumoniae.
Inducible gene expression systems are very useful to analyze cellular processes. The ability to switch the expression state of genes of interest may even be crucial if essential traits or genetic instability are involved. An integrative plasmid, pTEX2, was designed using the (anhydro)tetracycline-inducible promoter Pxyl/tet from staphylococcal plasmid pRAB11 to control gene expression in Streptococcus pneumoniae. The system was evaluated by expressing genes of the two-component regulatory system ciaRH of S. pneumoniae. With full induction of Pxyl/tet, wild-type levels of the response regulator CiaR were obtained, while the uninduced basal expression was low. Hyperactive variants of the kinase gene ciaH normally causing pronounced genetic instability could be handled without any problems upon cloning into pTEX2. Therefore, the expression system is well suited to express physiological levels of proteins in S. pneumoniae and also to aid regulatory studies.
Department of Microbiology, University of Kaiserslautern, D-67663 Kaiserslautern, Germany.
Marina
M
Meiers
Department of Microbiology, University of Kaiserslautern, D-67663 Kaiserslautern, Germany.
Anke
A
Laux
Department of Microbiology, University of Kaiserslautern, D-67663 Kaiserslautern, Germany.
Daniela
D
Eichinger
Department of Microbiology, University of Kaiserslautern, D-67663 Kaiserslautern, Germany.
Anne
A
Sexauer
Department of Microbiology, University of Kaiserslautern, D-67663 Kaiserslautern, Germany.
Patrick
P
Marx
Department of Microbial Genetics, Institute of Microbiology and Infection Medicine (IMIT), University of Tübingen, D-72076 Tübingen, Germany.|Research Department, Klinikum Nürnberg Medical School, Paracelsus Medical University, 90419 Nürnberg, Germany.
Ralph
R
Bertram
Department of Microbiology, University of Kaiserslautern, D-67663 Kaiserslautern, Germany.
Reinhold
R
Brückner
D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000235:genetics; D003001:Cloning, Molecular; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects; D010957:Plasmids; D011401:Promoter Regions, Genetic; D018085:Regulon; D015398:Signal Transduction; D013296:Streptococcus pneumoniae / Q000187:drug effects* / Q000235:genetics*; D013752:Tetracycline / Q000494:pharmacology*
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D013752:Tetracycline
Streptococcus pneumoniae; integrative plasmid; tetracycline-inducible expression; two-component regulatory system CiaRH