Genetic Manipulation and Virulence Assessment of Fusobacterium nucleatum. Issue 1 (15th June 2020)

• Record Type: Journal Article
• Authors: Peluso, Emily A.; Scheible, Matthew; Ton‐That, Hung; Wu, Chenggang
• Editors: Coico, Richard; McBride, Alison; Quarles, John M.; Stevenson, Brian; Taylor, Ronald K.
• Is Part Of: Current protocols in microbiology. Volume 57:Issue 1(2020)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2020-06-15
• Subjects: Fusobacterium nucleatum -- gene deletion -- infection model -- preterm birth -- transposon mutagenesis; Microbiology -- Laboratory manuals; Microbiology -- Technique -- Periodicals; Microbiological Techniques -- methods; Containment of Biohazards -- methods; Laboratory Infection -- prevention & control; Microbiology; Microbiology -- Technique; Fulltext; Internet Resources; Laboratory Manuals; Reference Materials; Periodicals; Laboratory manuals; 579.028
• DOI: 10.1002/cpmc.104
• Languages: English
• ISSNs: 1934-8525

Abstract

Abstract: Considered a commensal, the Gram‐negative anaerobe Fusobacterium nucleatum is a key member of the oral microbiome due to its wide range of interactions with many oral microbes. While the periodontal pathogenic properties of this organism have widely been examined, its connotation with extra‐oral infections, including preterm birth and colorectal cancer, has now become apparent. Nonetheless, little is known about the mechanisms of pathogenicity and the associated virulence factors of F. nucleatum, most likely due to limited genetic tools and facile methodology. Here, we describe molecular techniques for the genetic manipulation of F. nucleatum, including markerless, nonpolar gene deletion, complementation, and Tn5 transposon mutagenesis. Further, we provide methodology to assess virulence potential of F. nucleatum using a mouse model of preterm birth. © 2020 Wiley Periodicals LLC. Basic Protocol 1 : Generation of a galK mutant strain Basic Protocol 2 : Complementation of a mutant strain Basic Protocol 3 : Tn5 transposon mutagenesis of F. nucleatum Basic Protocol 4 : Mouse model of preterm birth