Archaeal Expression System (Sulfolobus shuttle vector)
Sulfolobus spp., a thermoacidophilic crenarchaeote, can be easily grown in
the laboratory and is interesting from a biotechnological point of view. Due
to their growth conditions (75-80°C and pH 3-3.5), enzymes from this organism
are thermostable and, if secreted, also acid-resistant and may have potential
for industrial applications. Furthermore,
Sulfolobus is developing into an important archaeal
model organism that is increasingly being
used in biochemical and molecular biology
So far, in vivo experiments in Sulfolobus were
hampered by the poor reproducibility of the few
described genetic systems. This technical hurdle
could be overcome with the developed Sulfolobus-
Escherichia coli shuttle vector.
Our construct meets the requirements for a small, episomal, easy-to-handle
multicopy vector. The different components are an E. coli replicon with a selectable
marker for E. coli, a Sulfolobus replicon, and a selectable marker for
Sulfolobus. The plasmid pRN1 was chosen as Sulfolobus replicon because of
its relatively small size of 5.4 kb. Our shuttle vector was found to be stable
in E. coli and to be suitable for cloning of additional DNA sequences. Furthermore,
the suitability of the vector constructs for reporter gene experiments
and for the expression of proteins in Sulfolobus was demonstrated.
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