Disruption of bbe02 by insertion of a luciferase gene increases transformation efficiency of Borrelia burgdorferi and allows live imaging in lyme disease susceptible C3H mice

Kamfai Chan, Laura Alter, Stephen W Barthold, Nikhat Parveen

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Lyme disease is the most prevalent tick-borne disease in North America and Europe. The causative agent, Borrelia burgdorferi persists in the white-footed mouse. Infection with B. burgdorferi can cause acute to persistent multisystemic Lyme disease in humans. Some disease manifestations are also exhibited in the mouse model of Lyme disease. Genetic manipulation of B. burgdorferi remains difficult. First, B. burgdorferi contains a large number of endogenous plasmids with unique sequences encoding unknown functions. The presence of these plasmids needs to be confirmed after each genetic manipulation. Second, the restriction modification defense systems, including that encoded by bbe02 gene lead to low transformation efficiency in B. burgdorferi. Therefore, studying the molecular basis of Lyme pathogenesis is a challenge. Furthermore, investigation of the role of a specific B. burgdorferi protein throughout infection requires a large number of mice, making it labor intensive and expensive. To overcome the problems associated with low transformation efficiency and to reduce the number of mice needed for experiments, we disrupted the bbe02 gene of a highly infectious and pathogenic B. burgdorferi strain, N40 D10/E9 through insertion of a firefly luciferase gene. The bbe02 mutant shows higher transformation efficiency and maintains luciferase activity throughout infection as detected by live imaging of mice. Infectivity and pathogenesis of this mutant were comparable to the wild-type N40 strain. This mutant will serve as an ideal parental strain to examine the roles of various B. burgdorferi proteins in Lyme pathogenesis in the mouse model in the future.

Original languageEnglish (US)
Article numbere0129532
JournalPLoS One
Volume10
Issue number6
DOIs
StatePublished - Jun 12 2015

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Borrelia burgdorferi
Lyme disease
Lyme Disease
Inbred C3H Mouse
luciferase
Luciferases
Genes
image analysis
Imaging techniques
mice
genes
Plasmids
pathogenesis
Firefly Luciferases
genetic engineering
mutants
plasmids
Infection
animal models
DNA Restriction-Modification Enzymes

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Disruption of bbe02 by insertion of a luciferase gene increases transformation efficiency of Borrelia burgdorferi and allows live imaging in lyme disease susceptible C3H mice. / Chan, Kamfai; Alter, Laura; Barthold, Stephen W; Parveen, Nikhat.

In: PLoS One, Vol. 10, No. 6, e0129532, 12.06.2015.

Research output: Contribution to journalArticle

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