title: Chemical and UV Mutagenesis.
abstract: The ability to create mutations is an important step towards understanding bacterial physiology and virulence. While targeted approaches are invaluable, the ability to produce genome-wide random mutations can lead to crucial discoveries. Transposon mutagenesis is a useful approach, but many interesting mutations can be missed by these insertions that interrupt coding and noncoding sequences due to the integration of an entire transposon. Chemical mutagenesis and UV-based random mutagenesis are alternate approaches to isolate mutations of interest with the potential of only single nucleotide changes. Once a standard method, difficulty in identifying mutation sites had decreased the popularity of this technique. However, thanks to the recent emergence of economical whole-genome sequencing, this approach to making mutations can once again become a viable option. Therefore, this chapter provides an overview protocol for random mutagenesis using UV light or DNA-damaging chemicals.
authors: Department of Microbiology, Molecular Genetics and Immunology, The University of Kansas Medical Center, MSN 3029, 3901 Rainbow Boulevard, Kansas City, KS, 66160, USA. jbose@kumc.edu.
authors: Jeffrey L
authors: JL
authors: Bose
mesh_terms: D004251:DNA Transposable Elements / Q000235:genetics*; D016680:Genome, Bacterial / Q000187:drug effects / Q000528:radiation effects; D008967:Molecular Biology / Q000379:methods*; D016296:Mutagenesis / Q000187:drug effects / Q000528:radiation effects; D009153:Mutagens / Q000633:toxicity; D009154:Mutation / Q000187:drug effects / Q000528:radiation effects; D009604:Nitrosoguanidines / Q000633:toxicity; D014466:Ultraviolet Rays / Q000009:adverse effects
chemical_list: D004251:DNA Transposable Elements; D009153:Mutagens; D009604:Nitrosoguanidines
keywords: Chemical mutagenesis; Mutation; Nitrosoguanidine; Random mutagenesis; UV mutagenesis