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  • 1
    ISSN: 1617-4623
    Keywords: Streptomyces azureus ; Thiostrepton resistance ; Tandem promoters ; Nuclease S1 mapping ; In vitro transcription
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Nuclease S1 protection experiments indicated that the thiostrepton resistance gene (tsr) of Streptomyces azureus is transcribed from tandem promoters, tsrp1 and tsrp2, that initiate transcription 45 and 173 nucleotides, respectively, upstream of the presumptive translational start codon. The −10 regions of both promoters show similarity to the consensus sequence for the major class of prokaryotic promoters, but the −35 regions do not, although they show some similarity to each other. Replacement of sequences upstream of position −22 relative to the tsrp2 start site with two different DNA segments affected the levels of the tsrp2 transcript but did not alter the tsrp2 initiation site. In vitro transcription assays using RNA polymerase from Streptomyces coelicolor A3(2) also confirmed the location of tsrp2 and identified additional start sites near tsrp2 that were barely detectable with in vivo synthesised RNA. Transcripts corresponding to initiation in vitro at tsrp1 could not be detected, suggesting that additional factors are required for utilisation of this promoter.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Molecular genetics and genomics 209 (1987), S. 101-109 
    ISSN: 1617-4623
    Keywords: Promoters ; S1 mapping ; In vitro transcription ; Protein secretion ; Signal peptide
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The DNA sequence of a 1.77 kb region of the Streptomyces coelicolor chromosome containing the coding and regulatory regions of the extracellular agarase (dagA) gene was determined. The sequence predicts a primary translation product of 309 amino acids and Mr 35132. Comparison of the N-terminal sequence determined for the mature extracellular protein with that of the primary translation product deduced from the DNA sequence predicts the presence of a 30 amino acid signal peptide. Analysis of the transcription of the dagA gene using high resolution S1 mapping, in vitro transcription, dinucleotide-primed in vitro transcription and in vivo promoter probing identified four promoters, initiating transcription approximately 32, 77, 125 and 220 nucleotides upstream of the coding sequence.
    Type of Medium: Electronic Resource
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