Development of a shuttle vector for Moraxella catarrhalis.
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.
Wang
Wei
W
Department of Microbiology, University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, TX 75390-9048, USA.
Attia
Ahmed S
AS
Liu
Lixia
L
Rosche
Thomas
T
Wagner
Nikki J
NJ
Hansen
Eric J
EJ
D018829:Adhesins, Bacterial / Q000235:genetics; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics*; D003001:Cloning, Molecular; D004269:DNA, Bacterial; D024881:Drug Resistance, Bacterial; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D006193:Haemophilus influenzae / Q000235:genetics; D001936:Moraxella catarrhalis / Q000235:genetics*; D009154:Mutation / Q000235:genetics; D010641:Phenotype; D010957:Plasmids / Q000235:genetics*; D018741:Replication Origin; D000198:Spectinomycin / Q000494:pharmacology; D014169:Transformation, Bacterial
D018829:Adhesins, Bacterial; D001425:Bacterial Outer Membrane Proteins; D004269:DNA, Bacterial; C089675:UspA protein, Moraxella catarrhalis; C045157:adhesin, Moraxella; D000198:Spectinomycin