Oligonucleotide-mediated genetic transformation of Borrelia burgdorferi

D. Scott Samuels, Claude F. Garon

Research output: Contribution to journalArticlepeer-review

Abstract

We have used short oligonucleotides to genetically transform the Lyme disease spirochaete Borrelia burgdorferi. The oligonucleotides are derived from the sequence of an Arg-133 to Ile mutant gyrB (chromosomal) gene that confers resistance to the antibiotic coumermycin A1. Oligonucleotides were about 10,000-fold less efficient at transformation, on a molar basis, than longer PCR-generated substrates. All of the transformants tested contained the predicted site-directed silent mutation in their gyrB genes. Antisense oligonucleotides were more efficient at transformation than either sense or double-stranded oligonucleotides. This is the first demonstration of oligonucleotides used to introduce site-directed mutations directly into the genome of a bacterium.

Original languageEnglish
Pages (from-to)519-522
Number of pages4
JournalMicrobiology
Volume143
Issue number2
DOIs
StatePublished - Feb 1997

Keywords

  • Borrelia burgdorferi
  • Electroporation
  • Genetic transformation
  • Lyme disease
  • Oligonucleotide
  • gyrB

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