TABLE 1.

Plasmids used for this study

PlasmidDescriptionPurpose of constructionReference or source
pMJ02SSV1 in pUC18 with lacS under control of tf55alpha promoterPromoter studies in Sulfolobus (heat shock induction)16
pMJ03pyrEF cassette inserted into pMJ02Selectable marker added for stability16
pMJ05pyrEF cassette, tf55alpha promoter, lacS from pBR05 inserted into pMJ02, replacing tf55alpha and lacS from pMJ02Destination vector for SulfolobusThis study
pMJ11Derivative of pMJ03, with NcoI site introduced to separate tf55alpha promoter from lacS; no peptide spacerPromoter studies in Sulfolobus (heat shock induction)S. Albers and M. Jonuscheit, unpublished
pBR-05pBR322 containing pyrEF cassette and lacS under control of tf55alpha promoter, with BssHII site introduced (Fig. 1)E. coli entry vector with heat shock promoterThis study
pASK5pASK75 containing sor gene fused to C-terminal Strep tagPCR template for amplification of Strep-tagged sor37
pBR-sorpBR05 containing sor gene fused to C-terminal Strep tagE. coli donor vector for sorThis study
pMJ05-sorpyrEF cassette, tf55alpha promoter, sor from pBR05-sor inserted into pMJ02, replacing tf55alpha and lacS from pMJ02sor expression in SulfolobusThis study
pSVA1pBR05 with an ApaI restriction site downstream of the stop codon of lacSE. coli entry vector with terminator sequence separatedThis study
pSVA2SSO2316-6his inserted into pSVA1 (Fig. 1)E. coli donor vector for SSO2316This study
pSVA5pSVA1 containing the lacS gene under control of araS promoter (Fig. 1)E. coli entry vector with ara promoterThis study
pSVA6tf55alpha promoter, SSO2316 from pSVA2 inserted into pMJ05, replacing tf55alpha and lacS from pMJ05SSO2316 expression in SulfolobusThis study
pSVA9araS promoter, lacS from pSVA5 inserted into pMJ05, replacing tf55-alpha promoter and lacS from pMJ05Promoter studies in Sulfolobus (arabinose induction)S. Albers, unpublished
pSVA14SSO2680-10his inserted into pSVA5E. coli donor vector for SSO2680This study
pSVA15araS promoter, SSO2680-10his from pSVA14 inserted into pMJ05, replacing tf55alpha and lacS from pMJ05SSO2680 expression in SulfolobusThis study
pSVA30SSO0287-Strep-10his inserted into pSVA5E. coli donor vector for SSO0287This study
pSVA31araS promoter, SSO0287-Strep-10his from pSVA30 inserted into pMJ05, replacing tf55alpha and lacS from pMJ05SSO0287 expression in SulfolobusThis study