Improved methods for genetic manipulation of the alkaliphile .

An improved approach was developed for the genetic manipulation of the Gram-positive extremophile (formerly called ). We describe an allelic replacement method originally developed for that allows the deletion, mutation, or insertion of genes without leaving markers or other genetic scars. In addition, a protocol for rapid plasmid methylation and transformation is presented. The combined methods allow the routine genetic manipulation of from initial transformation to the desired strain in 8 days. These methods improve as a model organism for the study of extremophiles.