Development of a LacZ-based Transcriptional Reporter System for Use with Moraxella Catarrhalis

Overview

Journal Plasmid

Date 2012 Dec 11

PMID 23219721

Citations 2

Authors

Amanda S Evans

Christine Pybus

Eric J Hansen

Affiliations

Soon will be listed here.

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.

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References

Hsieh Y, Wanner B . Global regulation by the seven-component Pi signaling system. Curr Opin Microbiol. 2010; 13(2):198-203. PMC: 2847643. DOI: 10.1016/j.mib.2010.01.014. View

Hays J, Eadie K, Verduin C, Verbrugh H, van Belkum A . A novel plasmid (pEMCJH03) isolated from moraxella catarrhalis possibly useful as a cloning and expression vector within this species. Plasmid. 2005; 53(3):263-8. DOI: 10.1016/j.plasmid.2004.11.003. View

Helminen M, Maciver I, Paris M, Latimer J, Lumbley S, Cope L . A mutation affecting expression of a major outer membrane protein of Moraxella catarrhalis alters serum resistance and survival in vivo. J Infect Dis. 1993; 168(5):1194-201. DOI: 10.1093/infdis/168.5.1194. View

Beaulieu D, Ouellette M, Bergeron M, Roy P . Characterization of a plasmid isolated from Branhamella catarrhalis and detection of plasmid sequences within the genome of a B. catarrhalis strain. Plasmid. 1988; 20(2):158-62. DOI: 10.1016/0147-619x(88)90020-0. View

Armbruster C, Hong W, Pang B, Weimer K, Juneau R, Turner J . Indirect pathogenicity of Haemophilus influenzae and Moraxella catarrhalis in polymicrobial otitis media occurs via interspecies quorum signaling. mBio. 2010; 1(3). PMC: 2925075. DOI: 10.1128/mBio.00102-10. View

Hoopman T, Liu W, Joslin S, Pybus C, Brautigam C, Hansen E . Identification of gene products involved in the oxidative stress response of Moraxella catarrhalis. Infect Immun. 2010; 79(2):745-55. PMC: 3028835. DOI: 10.1128/IAI.01060-10. View

Wang W, Kinkel T, Martens-Habbena W, Stahl D, Fang F, Hansen E . The Moraxella catarrhalis nitric oxide reductase is essential for nitric oxide detoxification. J Bacteriol. 2011; 193(11):2804-13. PMC: 3133116. DOI: 10.1128/JB.00139-11. View

Wang W, Hansen E . Plasmid pWW115, a cloning vector for use with Moraxella catarrhalis. Plasmid. 2006; 56(2):133-7. DOI: 10.1016/j.plasmid.2006.03.002. View

Furano K, Campagnari A . Inactivation of the Moraxella catarrhalis 7169 ferric uptake regulator increases susceptibility to the bactericidal activity of normal human sera. Infect Immun. 2003; 71(4):1843-8. PMC: 152102. DOI: 10.1128/IAI.71.4.1843-1848.2003. View

10.

Wang W, Attia A, Liu L, Rosche T, Wagner N, Hansen E . Development of a shuttle vector for Moraxella catarrhalis. Plasmid. 2005; 55(1):50-7. DOI: 10.1016/j.plasmid.2005.07.004. View

11.

Nguyen T, Zhang Z, Huang I, Wu C, Merritt J, Shi W . Genes involved in the repression of mutacin I production in Streptococcus mutans. Microbiology (Reading). 2009; 155(Pt 2):551-556. PMC: 2946218. DOI: 10.1099/mic.0.021303-0. View

12.

JUNI E, Heym G, Avery M . Defined medium for Moraxella (Branhamella) catarrhalis. Appl Environ Microbiol. 1986; 52(3):546-51. PMC: 203570. DOI: 10.1128/aem.52.3.546-551.1986. View

13.

Hoopman T, Liu W, Joslin S, Pybus C, Sedillo J, Labandeira-Rey M . Use of the chinchilla model for nasopharyngeal colonization to study gene expression by Moraxella catarrhalis. Infect Immun. 2011; 80(3):982-95. PMC: 3294655. DOI: 10.1128/IAI.05918-11. View

14.

Karalus R, Campagnari A . Moraxella catarrhalis: a review of an important human mucosal pathogen. Microbes Infect. 2000; 2(5):547-59. DOI: 10.1016/s1286-4579(00)00314-2. View

15.

Davie J, Earl J, de Vries S, Ahmed A, Hu F, Bootsma H . Comparative analysis and supragenome modeling of twelve Moraxella catarrhalis clinical isolates. BMC Genomics. 2011; 12:70. PMC: 3045334. DOI: 10.1186/1471-2164-12-70. View

16.

Murphy T, Brauer A, Grant B, Sethi S . Moraxella catarrhalis in chronic obstructive pulmonary disease: burden of disease and immune response. Am J Respir Crit Care Med. 2005; 172(2):195-9. PMC: 2718466. DOI: 10.1164/rccm.200412-1747OC. View

17.

Zhang X, Bremer H . Control of the Escherichia coli rrnB P1 promoter strength by ppGpp. J Biol Chem. 1995; 270(19):11181-9. DOI: 10.1074/jbc.270.19.11181. View

18.

Simons R, Houman F, Kleckner N . Improved single and multicopy lac-based cloning vectors for protein and operon fusions. Gene. 1987; 53(1):85-96. DOI: 10.1016/0378-1119(87)90095-3. View

19.

Wang W, Richardson A, Martens-Habbena W, Stahl D, Fang F, Hansen E . Identification of a repressor of a truncated denitrification pathway in Moraxella catarrhalis. J Bacteriol. 2008; 190(23):7762-72. PMC: 2583601. DOI: 10.1128/JB.01032-08. View

20.

Luke N, Jurcisek J, Bakaletz L, Campagnari A . Contribution of Moraxella catarrhalis type IV pili to nasopharyngeal colonization and biofilm formation. Infect Immun. 2007; 75(12):5559-64. PMC: 2168369. DOI: 10.1128/IAI.00946-07. View