A transcriptional reporter for in vivo promoter analysis in the archaeon Haloferax volcanii.
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.
Palmer
J R
JR
Department of Microbiology, Ohio State University, Columbus 43210.
Daniels
C J
CJ
D001483:Base Sequence; D004269:DNA, Bacterial / Q000235:genetics; D005798:Genes, Bacterial; D017930:Genes, Reporter; D016931:Halobacteriales / Q000235:genetics*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D012361:RNA, Transfer, Pro / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics; D014158:Transcription, Genetic
D004269:DNA, Bacterial; D012361:RNA, Transfer, Pro