ArticleApplied and environmental microbiology2012
A synthetic arabinose-inducible promoter confers high levels of recombinant protein expression in hyperthermophilic archaeon Sulfolobus islandicus.
Article in Applied and environmental microbiology, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 69 papers.
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69 citing papers in PubMed, 116 citations in OpenAlex.
- Cran1, member of a new class of OLD family ATPases, functions in cell cycle progression in an archaeon.EMBO reports · 2026Article
- An Lrs14 family protein functions as a nucleoid-associated protein regulating cell cycle progression in Sulfolobales.Communications biology · 2025Article
- Insights into the spool-like architecture and infection strategy of an enveloped archaeal virus.Science advances · 2025Article
- An archaeal chronic virus escapes the immunity of prokaryotic Argonaute.Nucleic acids research · 2025Article
- Archaeal replicative primase mediates DNA double-strand break repair.Nucleic acids research · 2025Article
- PerR functions as a redox-sensing transcription factor regulating metal homeostasis in the thermoacidophilic archaeon Saccharolobus islandicus REY15A.Nucleic acids research · 2025Article
- Regulatory sequence-based discovery of anti-defense genes in archaeal viruses.Nature communications · 2024Article
- Flexible TAM requirement of TnpB enables efficient single-nucleotide editing with expanded targeting scope.Nature communications · 2024Article
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- 5'-untranslated region sequences enhance plasmid-based protein production inFrontiers in microbiology · 2024Article
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- A clade of RHH proteins ubiquitous in Sulfolobales and their viruses regulates cell cycle progression.Nucleic acids research · 2023Article
- Dissection of Functional Domains of Orc1-2, the Archaeal Global DNA Damage-Responsive Regulator.International journal of molecular sciences · 2022Article
- DNA Motifs and an Accessory CRISPR Factor Determine Cas1 Binding and Integration Activity inInternational journal of molecular sciences · 2022Article
- Inactivation of Target RNA Cleavage of a III-B CRISPR-Cas System Induces Robust Autoimmunity inInternational journal of molecular sciences · 2022Article
- A Well-Conserved Archaeal B-Family Polymerase Functions as an Extender in Translesion Synthesis.mBio · 2022Article
- Progress and Challenges in Archaeal Genetic Manipulation.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Cyclic Tetra-Adenylate (cABiomolecules · 2021Article
- The biology of thermoacidophilic archaea from the order Sulfolobales.FEMS microbiology reviews · 2021Article
9 more citing papers are in PubMed but not listed here.
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Authors and funding
8 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Despite major progresses in genetic studies of hyperthermophilic archaea, recombinant protein production in these organisms always suffers from low yields and a robust expression system is still in great demand. Here we report a versatile vector that confers high levels of protein expression in Sulfolobus islandicus, a hyperthermophilic crenarchaeon. Two expression vectors, pSeSD and pEXA, harboring 11 unique restriction sites were constructed. They contain coding sequences of two hexahistidine (6×His) peptide tags and those coding for two protease sites, the latter of which make it possible to remove the peptide tags from expressed recombinant proteins. While pEXA employed an araS promoter for protein expression, pSeSD utilized P(araS-SD), an araS derivative promoter carrying an engineered ribosome-binding site (RBS; a Shine-Dalgarno [SD] sequence). We found that P(araS-SD) directed high levels of target gene expression. More strikingly, N-terminal amino acid sequencing of recombinant proteins unraveled that the protein synthesized from pEXA-N-lacS lacked the designed 6×His tag and that translation initiation did not start at the ATG codon of the fusion gene. Instead, it started at multiple sites downstream of the 6×His codons. Intriguingly, inserting an RBS site upstream of the ATG codon regained the expression of the 6×His tag, as shown with pSeSD-N-lacS. These results have yielded novel insight into the archaeal translation mechanism. The crenarchaeon Sulfolobus can utilize N-terminal coding sequences of proteins to specify translation initiation in the absence of an RBS site.
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