REPLACR-mutagenesis, a one-step method for site-directed mutagenesis by recombineering.

ABSTRACT: Mutagenesis is an important tool to study gene regulation, model disease-causing mutations and for functional characterisation of proteins. Most of the current methods for mutagenesis involve multiple step procedures. One of the most accurate methods for genetically altering DNA is recombineering, which uses bacteria expressing viral recombination proteins. Recently, the use of in vitro seamless assembly systems using purified enzymes for multiple-fragment cloning as well as mutagenesis is gaining ground. Although these in vitro isothermal reactions are useful when cloning multiple fragments, for site-directed mutagenesis it is unnecessary. Moreover, the use of purified enzymes in vitro is not only expensive but also more inaccurate than the high-fidelity recombination inside bacteria. Her

SUBMITTER: Trehan A 

PROVIDER: S-EPMC4707547 | biostudies-literature | 2016 Jan

REPOSITORIES: biostudies-literature