Conditions for gene disruption by homologous recombination of exogenous DNA into theSulfolobus solfataricusgenome

Abstract

The construction of directed gene deletion mutants is an essential tool in molecular biology that allows functional studies on the role of genes in their natural environment. For hyperthermophilic archaea, it has been difficult to obtain a reliable system to construct such mutants. However, during the past years, systems have been developed forThermococcus kodakarensisand twoSulfolobusspecies,S. acidocaldariusand derivatives ofS. solfataricus98/2. Here we describe an optimization of the method for integration of exogenous DNA intoS. solfataricusPBL 2025, anS. solfataricus98/2 derivative, based on lactose auxotrophy that now allows for routine gene inactivation.