| JWissmille 2004-10-23, 7:08 pm |
| http://www.ncbi.nlm.nih.gov/entrez/...db=pubmed&dopt=
Abstract&list_uids=15466540
* *Appl Environ Microbiol. 2004 Oct;70(10):5973-9.
*Genome-Wide Transposon Mutagenesis of Borrelia burgdorferi for
Identification of Phenotypic Mutants.*
*Stewart PE, Hoff J, Fischer E, Krum JG, Rosa PA.*
Rocky Mountain Laboratories, NIAID, NIH, 903 South 4th St., Hamilton, MT
59840. pestewart@niaid.nih.gov.
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.
PMID: 15466540 [PubMed - in process]
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