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

Appl Environ Microbiol. 1994 Oct;60(10):3867-9. doi: 10.1128/aem.60.10.3867-3869.1994.

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.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Base Sequence
  • DNA, Bacterial / genetics
  • Genes, Bacterial
  • Genes, Reporter*
  • Halobacteriales / genetics*
  • Molecular Sequence Data
  • Promoter Regions, Genetic*
  • RNA, Transfer, Pro / genetics
  • Saccharomyces cerevisiae / genetics
  • Transcription, Genetic

Substances

  • DNA, Bacterial
  • RNA, Transfer, Pro