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Free keywords:
lactose auxotrophy, selection
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 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.
