The expression in thermophilic Archaea such as Sulfolobus solfataricus of genes encoding proteins of biotechnological relevance is an interesting goal to improve and expand industrial applications, but requires knowledge of basic physiological features and hence suitable genetic tools. Sulfolobus solfataricus, a thermoacidophilic aerobe archaeon, was chosen as a model host system for the study of gene transfer and regulation because it has been proven to be a versatile host for viruses and plasmids and its genome sequencing is in good progress. An E.coli/Sulfolobus shuttle vector, the pEXSs, was obtained and demonstrated to replicate autonomously and to express a thermoadapted hygromicin phosphotransferase as the selective marker (1). In order to drive a heterologous expression in S. solfataricus, the region upstream of the alcohol dehydrogenase (adh) gene was studied in vivo and proven to be responsible for the highly regulated expression of its gene (2). Studies are underway in order to better define the transcriptionally active regulatory sequences and transcription factors, as well as their interaction. Furthermore, an expression cassette containing the adh promoter region and the alcohol dehydrogenase coding sequence (adh-hT) from Bacillus stearothermophilus was cloned in the pEXSs vector; the presence of an active and specific bacillar ADH was detected in crude extracts of the HYG resistant clones by ADH-hT specific enzyme assay.
The ADH promoter region: a model system for gene regulation and heterologous expression in Sulfolobus solfataricus / Fiorentino, Gabriella; P., Contursi; R., Cannio; M., Rossi; Bartolucci, Simonetta. - STAMPA. - (1999), pp. ---. (Intervento presentato al convegno Third Meeting on Extremophiles as Cell Factories tenutosi a Graz, Austria nel June, 3-6).
The ADH promoter region: a model system for gene regulation and heterologous expression in Sulfolobus solfataricus
FIORENTINO, GABRIELLA;P. Contursi;BARTOLUCCI, SIMONETTA
1999
Abstract
The expression in thermophilic Archaea such as Sulfolobus solfataricus of genes encoding proteins of biotechnological relevance is an interesting goal to improve and expand industrial applications, but requires knowledge of basic physiological features and hence suitable genetic tools. Sulfolobus solfataricus, a thermoacidophilic aerobe archaeon, was chosen as a model host system for the study of gene transfer and regulation because it has been proven to be a versatile host for viruses and plasmids and its genome sequencing is in good progress. An E.coli/Sulfolobus shuttle vector, the pEXSs, was obtained and demonstrated to replicate autonomously and to express a thermoadapted hygromicin phosphotransferase as the selective marker (1). In order to drive a heterologous expression in S. solfataricus, the region upstream of the alcohol dehydrogenase (adh) gene was studied in vivo and proven to be responsible for the highly regulated expression of its gene (2). Studies are underway in order to better define the transcriptionally active regulatory sequences and transcription factors, as well as their interaction. Furthermore, an expression cassette containing the adh promoter region and the alcohol dehydrogenase coding sequence (adh-hT) from Bacillus stearothermophilus was cloned in the pEXSs vector; the presence of an active and specific bacillar ADH was detected in crude extracts of the HYG resistant clones by ADH-hT specific enzyme assay.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
