Open AccessGenetic Transformation of the Relapsing Fever Spirochete Borrelia hermsii: Stable Integration and Expression of Green Fluorescent Protein from Linear Plasmid 200
Author(s) -
Lindy M. Fine,
Christopher G. Earnhart,
Richard T. Marconi
Publication year - 2011
Publication title -
journal of bacteriology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.652
H-Index - 246
eISSN - 1067-8832
pISSN - 0021-9193
DOI - 10.1128/jb.05037-11
Subject(s) - biology , relapsing fever , borrelia , plasmid , virology , virulence , microbiology and biotechnology , green fluorescent protein , genetics , gene , borrelia burgdorferi , antibody
Tick-borne relapsing fever (TBRF) is a spirochetal disease caused by at least 15 differentBorrelia species. It is a serious human health concern in regions of endemicity throughout the world. Transmission to humans occurs through the bites of infectedOrnithodoros ticks. In North America, the primaryBorrelia species associated with human disease areB. hermsii andB. turicatae . Direct demonstration of the role of putative TBRF spirochete virulence factors in the disease process has been hindered by the lack of a genetic manipulation system and complete genome sequences. Expanding on recent developments in these areas, here we demonstrate the successful generation of a clone ofB. hermsii YOR that constitutively produces green fluorescent protein (GFP) (B. hermsii YOR::kan gfp ). This strain was generated through introduction of akan-gfp cassette into a noncoding region of the 200-kbB. hermsii linear plasmid lp200. Genetic manipulation did not affect the growth rate or trigger the loss of native plasmids.B. hermsii YOR::kan gfp retained infectivity and elicited host seroconversion. Stable production of GFP was demonstrated bothin vitro andin vivo . This study represents a significant step forward in the development of tools that can be employed to study the virulence mechanisms of TBRF spirochetes.