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 language | English |
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Pages (from-to) | 519-522 |
Number of pages | 4 |
Journal | Microbiology |
Volume | 143 |
Issue number | 2 |
DOIs | |
State | Published - Feb 1997 |
Keywords
- Borrelia burgdorferi
- Electroporation
- Genetic transformation
- Lyme disease
- Oligonucleotide
- gyrB