Protocol

In Vitro Mutagenesis Protocols

Volume 634 of the series Methods in Molecular Biology pp 411-419

Date:

Random Mutagenesis Using a Mutator Strain

  • Ghazala MuteebAffiliated withLaboratory of Transcription Biology, Center for DNA Fingerprinting and Diagnostics
  • , Ranjan SenAffiliated withLaboratory of Transcription Biology, Center for DNA Fingerprinting and Diagnostics Email author 

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Abstract

Random mutagenesis is a useful technique to study the functions of different gene products. Propagation of the genes cloned in plasmids through a mutator strain, like Escherichia coli XL1-red, produces randomly mutagenized plasmid libraries. This method offers a very simple and economic way of introducing random point mutations throughout the gene with a fairly high mutation rate. The whole process involves transformation and propagation of a plasmid containing the desired gene into the XL-1 red strain, isolating the mutagenized plasmid library and transforming this library into a desired strain for screening the mutant phenotype.

Key words

XL1-red™ Random mutation Mutagenized library E. coli Spontaneous mutation DNA repair