title: Development of a LacZ-based transcriptional reporter system for use with Moraxella catarrhalis.
abstract: The lack of a transcriptional reporter system for use in Moraxella catarrhalis has hindered studies of gene regulation in this pathogen. PCR and recombinant DNA methods were used to insert a multicloning site (MCS) and promoterless full-length Escherichia coli lacZ gene, flanked by transcriptional terminators both immediately upstream and downstream, into the M. catarrhalis recombinant plasmid pWW115. Insertion into the MCS in the newly constructed plasmid pASE222 of M. catarrhalis promoter regions controlled by either a repressor (i.e., NsrR) or activator (i.e., PhoB) yielded transcriptional fusion constructs that were appropriately responsive to signal inputs dependent on the host strain genotype, as measured quantitatively by means of a Miller β-galactosidase assay. The transcriptional reporter plasmid pASE222 should prove to be a useful tool for rapid screening of factors affecting gene expression in M. catarrhalis.
authors.affiliation: Department of Pediatrics, University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, TX 75390, USA. Amanda.Evans@UTSouthwestern.edu
authors.forename: Amanda S
authors.initials: AS
authors.lastname: Evans
authors.forename: Christine
authors.initials: C
authors.lastname: Pybus
authors.forename: Eric J
authors.initials: EJ
authors.lastname: Hansen
mesh_terms: D001483:Base Sequence; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D017930:Genes, Reporter; D007763:Lac Operon / Q000235:genetics*; D008969:Molecular Sequence Data; D001936:Moraxella catarrhalis / Q000187:drug effects / Q000235:genetics*; D010710:Phosphates / Q000494:pharmacology; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D014158:Transcription, Genetic / Q000187:drug effects
chemical_list: D010710:Phosphates