Development of a shuttle vector for Moraxella catarrhalis

Wei Wang, Ahmed S. Attia, Lixia Liu, Thomas Rosche, Nikki J. Wagner, Eric J. Hansen

Research output: Contribution to journalArticle

9 Scopus citations

Abstract

Efforts to perform genetic analysis in Moraxella catarrhalis have been hampered by the lack of a cloning vector. M. catarrhalis strain E22 was previously shown to contain plasmid pLQ510 which lacked a selectable antibiotic resistance marker. Several methods were used to eliminate unnecessary DNA from pLQ510. Then, a 1.2 kb spectinomycin resistance cartridge, a multiple cloning site, and the origin of replication from pACYC184 were cloned into this plasmid backbone to obtain the 7.2 kb plasmid pWW102B. This new plasmid could replicate in M. catarrhalis as well as in both Escherichia coli and Haemophilus influenzae. This shuttle vector was used to clone and express two different M. catarrhalis genes, respectively, encoding an adhesin and a protein involved in serum resistance. When these two plasmids were introduced into appropriate M. catarrhalis mutants, they complemented the phenotypic deficiency of each mutant. This is the first report of functional complementation in trans in this pathogen.

Original languageEnglish (US)
Pages (from-to)50-57
Number of pages8
JournalPlasmid
Volume55
Issue number1
DOIs
StatePublished - Jan 1 2006

Keywords

  • Complementation
  • Moraxella catarrhalis
  • Plasmid
  • Shuttle vector

ASJC Scopus subject areas

  • Molecular Biology

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  • Cite this

    Wang, W., Attia, A. S., Liu, L., Rosche, T., Wagner, N. J., & Hansen, E. J. (2006). Development of a shuttle vector for Moraxella catarrhalis. Plasmid, 55(1), 50-57. https://doi.org/10.1016/j.plasmid.2005.07.004