Development of a LacZ-based transcriptional reporter system for use with Moraxella catarrhalis

Amanda S. Evans, Christine Pybus, Eric J. Hansen

Research output: Contribution to journalArticle

2 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)180-185
Number of pages6
JournalPlasmid
Volume69
Issue number2
DOIs
Publication statusPublished - Mar 2013

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Keywords

  • Moraxella catarrhalis
  • Plasmid
  • Transcriptional reporter

ASJC Scopus subject areas

  • Molecular Biology

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