RT Journal Article SR Electronic(1) A1 Guiral, Sébastien A1 Hénard, Vincent A1 Laaberki, Maria-Halima A1 Granadel, Chantal A1 Prudhomme, Marc A1 Martin, Bernard A1 Claverys, Jean-PierreYR 2006 T1 Construction and evaluation of a chromosomal expression platform (CEP) for ectopic, maltose-driven gene expression in Streptococcus pneumoniae JF Microbiology, VO 152 IS 2 SP 343 OP 349 DO https://doi.org/10.1099/mic.0.28433-0 PB Microbiology Society, SN 1465-2080, AB In this paper, the construction and evaluation of a chromosomal expression platform (CEP), which allows controlled gene expression following ectopic integration into the chromosome of Streptococcus pneumoniae, is described. CEP is based on the well-studied maltosaccharide-inducible system. To facilitate integration at CEP, a plasmid, pCEP, capable of replication in Escherichia coli, but not in S. pneumoniae, was assembled. This plasmid contains an expression/selection cassette flanked on each side by more than 2 kb of pneumococcal DNA. The cassette comprises a maltose-inducible promoter, PM, separated from a kanamycin-resistance gene by NcoI and BamHI cloning sites. Clones harbouring the gene of interest integrated at CEP under the control of PM can be obtained through direct transformation of an S. pneumoniae recipient with ligation products between that gene and NcoI/BamHI-digested pCEP DNA, followed by selection for kanamycin-resistant transformants., UL https://www.microbiologyresearch.org/content/journal/micro/10.1099/mic.0.28433-0