A tetracycline-inducible integrative expression system for Streptococcus pneumoniae

作者：Meiers Marina; Laux Anke; Eichinger Daniela; Sexauer Anne; Marx Patrick; Bertram Ralph; Brueckner Reinhold

来源：FEMS Microbiology Letters, 2017, 364(5): fnx044.

DOI：10.1093/femsle/fnx044

• Streptococcus pneumoniae
• tetracycline-inducible expression
• integrative plasmid
• two-component regulatory system CiaRH

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 P-xyl/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 P-xyl/tet, wild-type levels of the response regulator CiaR were obtained, while theuninducedbasal 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.

• 出版日期2017-3