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. 1994 Oct;60(10):3867–3869. doi: 10.1128/aem.60.10.3867-3869.1994

A transcriptional reporter for in vivo promoter analysis in the archaeon Haloferax volcanii.

J R Palmer 1, C J Daniels 1
PMCID: PMC201898  PMID: 7986055

Abstract

We have used a modified intron-containing tRNA(Pro(UGG) gene (tRNA(ProM), derived from the Saccharomyces cerevisiae tRNA(Pro(UGG) gene, as a reporter to measure in vivo transcription from a halophilic archaeon promoter. Coupling of the yeast tRNA(ProM) gene to the Haloferax volcanii tRNA(Lys) promoter on the H. volcanii plasmid pWL201 led to the production of a single stable transcript that was readily quantitated by Northern (RNA) blot analysis. Comparison of tRNA(ProM) RNA production from constructs containing the wild-type tRNA(Lys) promoter and those containing mutant tRNA(Lys) promoters demonstrated that this assay system can be used to measure expression from strong and weak promoters.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Charlebois R. L., Lam W. L., Cline S. W., Doolittle W. F. Characterization of pHV2 from Halobacterium volcanii and its use in demonstrating transformation of an archaebacterium. Proc Natl Acad Sci U S A. 1987 Dec;84(23):8530–8534. doi: 10.1073/pnas.84.23.8530. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Cline S. W., Lam W. L., Charlebois R. L., Schalkwyk L. C., Doolittle W. F. Transformation methods for halophilic archaebacteria. Can J Microbiol. 1989 Jan;35(1):148–152. doi: 10.1139/m89-022. [DOI] [PubMed] [Google Scholar]
  3. Creti R., Londei P., Cammarano P. Complete nucleotide sequence of an archaeal (Pyrococcus woesei) gene encoding a homolog of eukaryotic transcription factor IIB (TFIIB). Nucleic Acids Res. 1993 Jun 25;21(12):2942–2942. doi: 10.1093/nar/21.12.2942. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Hain J., Reiter W. D., Hüdepohl U., Zillig W. Elements of an archaeal promoter defined by mutational analysis. Nucleic Acids Res. 1992 Oct 25;20(20):5423–5428. doi: 10.1093/nar/20.20.5423. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Hausner W., Frey G., Thomm M. Control regions of an archaeal gene. A TATA box and an initiator element promote cell-free transcription of the tRNA(Val) gene of Methanococcus vannielii. J Mol Biol. 1991 Dec 5;222(3):495–508. doi: 10.1016/0022-2836(91)90492-o. [DOI] [PubMed] [Google Scholar]
  6. Holmes M. L., Dyall-Smith M. L. Mutations in DNA gyrase result in novobiocin resistance in halophilic archaebacteria. J Bacteriol. 1991 Jan;173(2):642–648. doi: 10.1128/jb.173.2.642-648.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Lam W. L., Doolittle W. F. Shuttle vectors for the archaebacterium Halobacterium volcanii. Proc Natl Acad Sci U S A. 1989 Jul;86(14):5478–5482. doi: 10.1073/pnas.86.14.5478. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Langer D., Zillig W. Putative tfIIs gene of Sulfolobus acidocaldarius encoding an archaeal transcription elongation factor is situated directly downstream of the gene for a small subunit of DNA-dependent RNA polymerase. Nucleic Acids Res. 1993 May 11;21(9):2251–2251. doi: 10.1093/nar/21.9.2251. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Nieuwlandt D. T., Daniels C. J. An expression vector for the archaebacterium Haloferax volcanii. J Bacteriol. 1990 Dec;172(12):7104–7110. doi: 10.1128/jb.172.12.7104-7110.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Nieuwlandt D. T., Haas E. S., Daniels C. J. The RNA component of RNase P from the archaebacterium Haloferax volcanii. J Biol Chem. 1991 Mar 25;266(9):5689–5695. [PubMed] [Google Scholar]
  11. Ouzounis C., Sander C. TFIIB, an evolutionary link between the transcription machineries of archaebacteria and eukaryotes. Cell. 1992 Oct 16;71(2):189–190. doi: 10.1016/0092-8674(92)90347-f. [DOI] [PubMed] [Google Scholar]
  12. Palmer J. R., Nieuwlandt D. T., Daniels C. J. Expression of a yeast intron-containing tRNA in the archaeon Haloferax volcanii. J Bacteriol. 1994 Jun;176(12):3820–3823. doi: 10.1128/jb.176.12.3820-3823.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ponce M. R., Micol J. L. PCR amplification of long DNA fragments. Nucleic Acids Res. 1992 Feb 11;20(3):623–623. doi: 10.1093/nar/20.3.623. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Reiter W. D., Hüdepohl U., Zillig W. Mutational analysis of an archaebacterial promoter: essential role of a TATA box for transcription efficiency and start-site selection in vitro. Proc Natl Acad Sci U S A. 1990 Dec;87(24):9509–9513. doi: 10.1073/pnas.87.24.9509. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Rowlands T., Baumann P., Jackson S. P. The TATA-binding protein: a general transcription factor in eukaryotes and archaebacteria. Science. 1994 May 27;264(5163):1326–1329. doi: 10.1126/science.8191287. [DOI] [PubMed] [Google Scholar]
  16. Zillig W., Palm P., Reiter W. D., Gropp F., Pühler G., Klenk H. P. Comparative evaluation of gene expression in archaebacteria. Eur J Biochem. 1988 May 2;173(3):473–482. doi: 10.1111/j.1432-1033.1988.tb14023.x. [DOI] [PubMed] [Google Scholar]

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