title: Fluorescent reporters for markerless genomic integration in Staphylococcus aureus.
abstract: We present integration vectors for Staphylococcus aureus encoding the fluorescent reporters mAmetrine, CFP, sGFP, YFP, mCherry and mKate. The expression is driven either from the sarA-P1 promoter or from any other promoter of choice. The reporter can be inserted markerless in the chromosome of a wide range of S. aureus strains. The integration site chosen does not disrupt any open reading frame, provides good expression, and has no detectable effect on the strains physiology. As an intermediate construct, we present a set of replicating plasmids containing the same fluorescent reporters. Also in these reporter plasmids the sarA-P1 promoter can be replaced by any other promoter of interest for expression studies. Cassettes from the replication plasmids can be readily swapped with the integration vector. With these constructs it becomes possible to monitor reporters of separate fluorescent wavelengths simultaneously.
authors.affiliation: Medical Microbiology, University Medical Center Utrecht, Utrecht, The Netherlands.
authors.forename: Nienke W M
authors.initials: NW
authors.lastname: de Jong
authors.affiliation: Medical Microbiology, University Medical Center Utrecht, Utrecht, The Netherlands.
authors.forename: Thijs
authors.initials: T
authors.lastname: van der Horst
authors.affiliation: Medical Microbiology, University Medical Center Utrecht, Utrecht, The Netherlands.
authors.forename: Jos A G
authors.initials: JA
authors.lastname: van Strijp
authors.affiliation: Medical Microbiology, University Medical Center Utrecht, Utrecht, The Netherlands.|Laboratory of Phytopathology, Wageningen University, Wageningen, The Netherlands.
authors.forename: Reindert
authors.initials: R
authors.lastname: Nijland
mesh_terms: D055879:Gene Knock-In Techniques / Q000379:methods*; D017930:Genes, Reporter; D005822:Genetic Vectors; D008164:Luminescent Proteins / Q000032:analysis* / Q000235:genetics; D010957:Plasmids; D013211:Staphylococcus aureus / Q000235:genetics*
chemical_list: D008164:Luminescent Proteins