Recombineering: Advanced-Advanced Bacterial Genetics

Recombineering is homologous recombination-mediated in vivo genetic engineering. The author was introduced to John Roth at University of Utah in 1934, when Roth was teaching Biology 314, which also could have been called Advanced Bacterial Genetics. This chapter provides a summary of one of the most recent breakthroughs in genetic research, recombination-mediated genetic engineering, aka “recombineering.” A crucial difference from bacterial encoded recombination systems is that recombineering does not depend on RecA function. In the age of recombineering, creating desired mutations has become much easier. The chapter presents a novel method to optimize oligonucleotide recombineering; the technique is powerful in that it can be used to mutate essential genes.