Adaptation of a luciferase gene reporter and lac expression system to Borrelia burgdorferi

Jon S. Blevins, Andrew T. Revel, Alexandra H. Smith, Gulnaz N. Bachlani, Michael V. Norgard

Research output: Contribution to journalArticle

48 Citations (Scopus)

Abstract

The development of new genetic systems for studying the complex regulatory events that occur within Borrelia burgdorferi is an important goal of contemporary Lyme disease research. Although recent advancements have been made in the genetic manipulation of B. burgdorferi, there still remains a paucity of basic molecular systems for assessing differential gene expression in this pathogen. Herein, we describe the adaptation of two powerful genetic tools for use in B. burgdorferi. The first is a Photinus pyralis firefly luciferase gene reporter that was codon optimized to enhance translation in B. burgdorferi. Using this modified reporter, we demonstrated an increase in luciferase expression when B. burgdorferi transformed with a shuttle vector encoding the outer surface protein C (OspC) promoter fused to the luciferase reporter was cultivated in the presence of fresh rabbit blood. The second is a lac operator/repressor system that was optimized to achieve the tightest degree of regulation. Using the aforementioned luciferase reporter, we assessed the kinetics and maximal level of isopropyl-β-D-thiogalactopyranoside (IPTG)-dependent gene expression. This lac-inducible expression system also was used to express the gene carried on Ip25 required for borrelial persistence in ticks (bptA). These advancements should be generally applicable for assessing further the regulation of other genes potentially involved in virulence expression by B. burgdorferi.

Original languageEnglish (US)
Pages (from-to)1501-1513
Number of pages13
JournalApplied and Environmental Microbiology
Volume73
Issue number5
DOIs
StatePublished - Mar 2007

Fingerprint

Borrelia burgdorferi
Lac Operon
luciferase
Luciferases
Reporter Genes
reporter genes
gene expression
gene
Lyme disease
tick
virulence
Photinus pyralis
persistence
pathogen
Fireflies
blood
Lac Repressors
Thiogalactosides
Firefly Luciferases
Gene Expression

ASJC Scopus subject areas

  • Environmental Science(all)
  • Biotechnology
  • Microbiology

Cite this

Adaptation of a luciferase gene reporter and lac expression system to Borrelia burgdorferi. / Blevins, Jon S.; Revel, Andrew T.; Smith, Alexandra H.; Bachlani, Gulnaz N.; Norgard, Michael V.

In: Applied and Environmental Microbiology, Vol. 73, No. 5, 03.2007, p. 1501-1513.

Research output: Contribution to journalArticle

Blevins, Jon S. ; Revel, Andrew T. ; Smith, Alexandra H. ; Bachlani, Gulnaz N. ; Norgard, Michael V. / Adaptation of a luciferase gene reporter and lac expression system to Borrelia burgdorferi. In: Applied and Environmental Microbiology. 2007 ; Vol. 73, No. 5. pp. 1501-1513.
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