AN INSERTION ELEMENT OF THE EXTREMELY THERMOPHILIC ARCHAEON SULFOLOBUS-SOLFATARICUS TRANSPOSES INTO THE ENDOGENOUS BETA-GALACTOSIDASE GENE

Three phenotypically stable mutants of the extremely thermophilic archaeon Sulfolobus solfataricus have been isolated by screening for beta-galactosidase negative colonies on plates with X-Gal (5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside). From one of these mutants an insertion element, designated ISC1217, was isolated and characterized. Sequence analysis of ISC1217 and of the regions adjacent to the insertion site in the beta-galactosidase gene revealed features typical of a transposable element: ISC1217 contained terminal inverted repeats and was flanked by a direct repeat of 6 bp. The 1147 bp sequence contained an open reading frame encoding a putative protein of 354 amino acid residues and, overlapping this, two smaller open reading frames on the opposite strand. There were approximately 8 copies of the insertion element in the S. solfataricus genome. ISC1217 did not cross-hybridize with DNA of other Sulfolobus species. All three independently isolated beta-galactosidase mutants of S. solfataricus arose by transposition of ISC1217 or a related element.