Elimination of channel-forming activity by insertional inactivation of the p66 gene in Borrelia burgdorferi.
P66 is a chromosomally encoded 66-kDa integral outer membrane protein of the Lyme disease agent Borrelia burgdorferi exhibiting channel-forming activity. Herein, we inactivated and subsequently complemented the p66 gene in the B31-A (WT) strain. The P66 protein was also inactivated in two other channel-forming protein mutant strains, P13-18 (Deltap13) and Deltabba01, and then compared with the channel-forming activities of wild-type and various p66 mutant strains. We further investigated the ion-selectivity of native, purified P66. In conclusion, we show that the porin activity of P66 is eliminated by insertional inactivation and that this activity can be rescued by gene complementation.
Pinne
Marija
M
Department of Molecular Biology, Umeå University, Sweden.
Thein
Marcus
M
Denker
Katrin
K
Benz
Roland
R
Coburn
Jenifer
J
Bergström
Sven
S
D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D025065:Borrelia burgdorferi / Q000235:genetics* / Q000378:metabolism; D005816:Genetic Complementation Test; D015151:Immunoblotting; D007473:Ion Channels / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D016254:Mutagenesis, Insertional; D009154:Mutation; D018272:Porins / Q000235:genetics* / Q000378:metabolism / Q000502:physiology
D001426:Bacterial Proteins; D007473:Ion Channels; D008565:Membrane Proteins; C107893:Oms66 protein, Borrelia; D018272:Porins