A Novel Mutagenesis Strategy to Inactivate the Gene Encoding IgAl Protease in Neisseria gonorrhoeae for Use in the' Human Challenge Model
Koymen, Hakan O.
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Immunoglobulin Al (IgAI) protease is believed to be a possible virulence factor of Neisseria gonorrhoeae. However, until this point it has been impossible to study the role of IgAl protease in vivo. To avoid the addition of any new antibiotic resistance markers, our laboratory had developed and refined a novel two step mutagenesis strategy which allows us to manipulate a specific gene. Through this method we have successfully constructed an isogenic mutant which is unable to produce a functional IgAI protease. Furthermore, we have characterized our mutant, and found that all traits, including lipooligosaccharide, pilin type, and opacity proteins, remain intact and unchanged. Since we have not affected the genome in any other way, and have not given the bacteria any antibiotic advantages, we are now prepared to test the mutant by experimental urethral infection in our human challenge model.