Genome-wide transposon mutagenesis of Borrelia burgdorferi for identification of phenotypic mutants. Appl Environ Microbiol 2004 Oct;70(10):5973-9
Date
10/07/2004Pubmed ID
15466540Pubmed Central ID
PMC522107DOI
10.1128/AEM.70.10.5973-5979.2004Scopus ID
2-s2.0-8144223362 (requires institutional sign-in at Scopus site) 79 CitationsAbstract
The spirochete Borrelia burgdorferi is the causative agent of Lyme disease, the leading vector-borne illness in the United States. Many of the genetic factors affecting spirochete morphology and physiology are unknown due to the limited genetic tools available and the large number of open reading frames with unknown functions. By adapting a mariner transposon to function in B. burgdorferi, we have developed a random mutagenesis system that tags the mutated locus for rapid identification. Transposition occurs at saturating levels in B. burgdorferi and appears to be random, targeting both linear and circular replicons. By combining the transposon system with a screen for factors affecting growth rate, mutations were readily identified in genes putatively involved in cell division and chemotaxis and a hypothetical open reading frame involved in outer membrane integrity. The successful adaptation of a mariner transposon to function in B. burgdorferi should aid in identifying virulence factors and novel gene products related to spirochete physiology.
Author List
Stewart PE, Hoff J, Fischer E, Krum JG, Rosa PAAuthor
Jessica Hoff PhD Assistant Professor in the Pathology department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
AnimalsBase Sequence
Borrelia burgdorferi
Cell Division
DNA Transposable Elements
DNA, Bacterial
Genome, Bacterial
Humans
Microscopy, Electron, Scanning
Mutagenesis, Insertional
Mutation
Open Reading Frames
Phenotype
Plasmids
Virulence
