ArticleApplied and environmental microbiology2008
Shuttle vector expression in Thermococcus kodakaraensis: contributions of cis elements to protein synthesis in a hyperthermophilic archaeon.
Article in Applied and environmental microbiology, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 66 papers.
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Who cites it
66 citing papers in PubMed, 111 citations in OpenAlex.
- CRISPR spacers reveal diverse and abundant Thermococcales viruses in hydrothermal vents.Research square · 2026Article
- Removal and bypass of restriction modification systems increases transformation efficiency ofFrontiers in microbiology · 2026Article
- The arginine synthetase pathway is responsible for carbamoyl phosphate synthesis inFrontiers in microbiology · 2026Article
- Pentose bisphosphate pathway can act in central metabolism for nucleoside-dependent growth ofApplied and environmental microbiology · 2025Article
- 5'-untranslated region sequences enhance plasmid-based protein production inFrontiers in microbiology · 2024Article
- A self-transmissible plasmid from a hyperthermophile that facilitates genetic modification of diverse Archaea.Nature microbiology · 2023Article
- Biochemical and genetic examination of two aminotransferases from the hyperthermophilic archaeonFrontiers in microbiology · 2023Article
- Progress and Challenges in Archaeal Genetic Manipulation.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Transformation Techniques for the Anaerobic Hyperthermophile Thermococcus kodakarensis.Methods in molecular biology (Clifton, N.J.) · 2022Article
- TK1211 Encodes an Amino Acid Racemase towards Leucine and Methionine in the Hyperthermophilic Archaeon Thermococcus kodakarensis.Journal of bacteriology · 2021Article
- High-efficiency transformation of archaea by direct PCR products with its application to directed evolution of a thermostable enzyme.Microbial biotechnology · 2021Article
- Thermococcus kodakarensis provides a versatile hyperthermophilic archaeal platform for protein expression.Methods in enzymology · 2021Article
- The Hyperthermophilic Restriction-Modification Systems ofFrontiers in microbiology · 2021Article
- A Structurally Novel Lipoyl Synthase in the Hyperthermophilic Archaeon Thermococcus kodakarensis.Applied and environmental microbiology · 2020Article
- Integration of large heterologous DNA fragments into the genome of Thermococcus kodakarensis.Extremophiles : life under extreme conditions · 2020Article
- An overview of 25 years of research on Thermococcus kodakarensis, a genetically versatile model organism for archaeal research.Folia microbiologica · 2020Review
- The global distribution and evolutionary history of the pT26-2 archaeal plasmid family.Environmental microbiology · 2019Article
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- TFS and Spt4/5 accelerate transcription through archaeal histone-based chromatin.Molecular microbiology · 2019Article
6 more citing papers are in PubMed but not listed here.
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
Abstract
Shuttle vectors that replicate stably and express selectable phenotypes in both Thermococcus kodakaraensis and Escherichia coli have been constructed. Plasmid pTN1 from Thermococcus nautilis was ligated to the commercial vector pCR2.1-TOPO, and selectable markers were added so that T. kodakaraensis transformants could be selected by DeltatrpE complementation and/or mevinolin resistance. Based on Western blot measurements, shuttle vector expression of RpoL-HA, a hemagglutinin (HA) epitope-tagged subunit of T. kodakaraensis RNA polymerase (RNAP), was approximately 8-fold higher than chromosome expression. An idealized ribosome binding sequence (5'-AGGTGG) was incorporated for RpoL-HA expression, and changes to this sequence reduced expression. Changing the translation initiation codon from AUG to GUG did not reduce RpoL-HA expression, but replacing AUG with UUG dramatically reduced RpoL-HA synthesis. When functioning as translation initiation codons, AUG, GUG, and UUG all directed the incorporation of methionine as the N-terminal residue of RpoL-HA synthesized in T. kodakaraensis. Affinity purification confirmed that an HA- plus six-histidine-tagged RpoL subunit (RpoL-HA-his(6)) synthesized ectopically from a shuttle vector was assembled in vivo into RNAP holoenzymes that were active and could be purified directly from T. kodakaraensis cell lysates by Ni(2+) binding and imidazole elution.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.