Conditions for gene disruption by homologous recombination of exogenous DNA into the Sulfolobus solfataricus genome.
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 for Thermococcus kodakarensis and two Sulfolobus species, S. acidocaldarius and derivatives of S. solfataricus 98/2. Here we describe an optimization of the method for integration of exogenous DNA into S. solfataricus PBL 2025, an S. solfataricus 98/2 derivative, based on lactose auxotrophy that now allows for routine gene inactivation.
Albers
Sonja-Verena
SV
Department of Microbiology, Groningen Biomolecular Sciences and Biotechnology Institute and the Zernike Institute for Advanced Materials, University of Groningen, Kerklaan 30, 9751 NN HAREN, The Netherlands. s.v.albers@rug.nl
Driessen
Arnold J M
AJ
D003470:Culture Media; D019641:DNA, Archaeal / Q000235:genetics*; D018274:Electroporation; D017353:Gene Deletion; D005821:Genetic Techniques; D020745:Genome, Archaeal; D007785:Lactose / Q000378:metabolism; D011995:Recombination, Genetic; D012641:Selection, Genetic; D048229:Sulfolobus solfataricus / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism
D003470:Culture Media; D019641:DNA, Archaeal; D007785:Lactose