Screening for novel antibiotic resistance genes

Methods Mol Biol. 2010:668:265-71. doi: 10.1007/978-1-60761-823-2_18.

Abstract

Knowledge of novel antibiotic resistance genes aids in the understanding of how antibiotics function and how bacteria fight them. This knowledge also allows future generations of an antibiotic or antibiotic group to be altered to allow the greatest efficacy. The method described here is very simple in theory. The bacterial strains are screened for antibiotic resistance. Cultures of the strain are grown, and DNA is extracted. A partial digest of the extraction is cloned into Escherichia coli, and the transformants are plated on selective media. Any colony that grows will possess the antibiotic resistance gene and can be further examined. In actual practice, however, this technique can be complicated. The detailed protocol will need to be optimized for each bacterial strain, vector, and cell line chosen.

MeSH terms

  • Animals
  • Anti-Bacterial Agents / pharmacology*
  • Cloning, Molecular
  • Drug Resistance, Microbial / genetics*
  • Escherichia coli* / drug effects
  • Escherichia coli* / genetics
  • Humans
  • Microbial Sensitivity Tests / instrumentation
  • Microbial Sensitivity Tests / methods
  • Sequence Analysis, DNA

Substances

  • Anti-Bacterial Agents