Allelic Exchange of the Flagellin Gene fliC of Clositridium difficile: Generation of a Knockout Mutant

Abstract

Clostridium difficile is an antibiotic-resistant Gram-positive bacterium. It has become a major concern for hospitals and acute care medical wards where antibiotics are used more readily. Therapies and preventative measures are being developed to control the disease, and therefore it is crucial to understand the basis of its colonization of susceptible hosts (e.g. antibiotic-treated patients). Genetic manipulation of the C. difficile genome can be achieved through allelic exchange technologies. Precise alterations are made in the PCR-ribotype 027 strain, R20291 to generate a deletion mutant of the flagellin gene fliC. By following mutagenesis protocols, this mutant can be obtained to serve as a tool and for studies on C. difficile motility and chemotaxis.