Inactivation of the flagellin gene flaA in Magnetospirillum gryphiswaldense results in nonmagnetotactic mutants lacking flagellar filaments.
Magnetotactic bacteria synthesize magnetosomes, which cause them to orient and migrate along magnetic field lines. The analysis of magnetotaxis and magnetosome biomineralization at the molecular level has been hindered by the unavailability of genetic methods, namely the lack of a means to introduce directed gene-specific mutations. Here we report a method for knockout mutagenesis by homologous recombination in Magnetospirillum gryphiswaldense. Multiple flagellin genes, which are unlinked in the genome, were identified in M. gryphiswaldense. The targeted disruption of the flagellin gene flaA was shown to eliminate flagella formation, motility, and magnetotaxis. The techniques described in this paper will make it possible to take full advantage of the forthcoming genome sequences of M. gryphiswaldense and other magnetotactic bacteria.
Schultheiss
Daniel
D
Max Planck Institute for Marine Microbiology, 28359 Bremen, Germany.
Kube
Michael
M
Schüler
Dirk
D
D003227:Conjugation, Genetic; D005407:Flagella / Q000378:metabolism*; D005408:Flagellin / Q000235:genetics* / Q000378:metabolism; D017353:Gene Deletion; D008280:Magnetics; D042201:Magnetospirillum / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D008969:Molecular Sequence Data; D009068:Movement; D010957:Plasmids; D011995:Recombination, Genetic
D005408:Flagellin; C067138:flaA protein, bacteria