Genetic elements for selection, deletion mutagenesis and complementation in Francisella spp.
Francisella novicida is a gram-negative pathogen that can induce disease in mice that mimics human tularemia, and is nearly identical to Francisella tularensis at the genomic level. In this work a number of antibiotic marker cassettes that incorporate a strong F. novicida promoter is constructed, which greatly enhances selection in F. novicida and F. tularensis. Two low-copy plasmid vectors based on a broad-host-range plasmid, and an integrating vector have also been made, and these can be used for genetic complementation. Two general approaches to deletion mutagenesis in F. novicida is also described.
Ludu
Jagjit S
JS
Department of Biochemistry and Microbiology, University of Victoria, Victoria, BC, Canada.
Nix
Eli B
EB
Duplantis
Barry N
BN
de Bruin
Olle M
OM
Gallagher
Larry A
LA
Hawley
Laura M
LM
Nano
Francis E
FE
D000818:Animals; D002642:Chick Embryo; D003001:Cloning, Molecular; D005603:Francisella / Q000235:genetics* / Q000472:pathogenicity; D005816:Genetic Complementation Test; D005822:Genetic Vectors / Q000235:genetics*; D016905:Gram-Negative Bacterial Infections / Q000382:microbiology; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010957:Plasmids; D011401:Promoter Regions, Genetic; D017384:Sequence Deletion; D014169:Transformation, Bacterial