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  • Glucose kinase  (1)
  • Nuclease S1 mapping  (1)
  • Recombinant DNA  (1)
  • 1
    Electronic Resource
    Electronic Resource
    The repressor gene (c) of the Streptomyces temperate phage ϕc31: Nucleotide sequence, analysis and functional cloning (1988)
    Springer
    Molecular genetics and genomics 213 (1988), S. 269-277 
    Details
    ISSN: 1617-4623
    Keywords: Recombinant DNA ; Helix-turn-helix motifs ; In vitro transcription-translation ; Phage immunity ; Exonuclease III deletions
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The nucleotide sequence of the 3.4 kb SphI-G fragment that contained the repressor gene (c) of the temperate Streptomyces phage ϕc31 was determined. Analysis of this sequence revealed a large open reading frame with protein coding character and sequence changes in c gene point and deletion mutants identified this as the coding region of the repressor. Two of the mutants studied had undergone deletions of 1.1 kb and 1.4 kb that had occurred across short direct repeats of 6 bp and 11 bp, respectively. Coupled in vitro transcription-translation experiments using the cloned SphI-G fragment and Streptomyces lividans cell free extracts identified a protein product of approximately 72 kDa, in close agreement with that predicted from the nucleotide sequence. A strongly predicted helix-turn-helix motif that may be involved in DNA binding occurred towards the carboxy-terminus of the amino acid sequence. Initial attempts to clone the SphI-G fragment in Streptomyces failed; using information gained from the sequence analysis a smaller segment of this DNA fragment was cloned in S. lividans and conferred immunity to a clear plaque mutant (c1) of ϕc31.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00339591
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    Paper (German National Licenses)
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  • 2
    Electronic Resource
    Electronic Resource
    Tandem promoters, tsrp1 and tsrp2, direct transcription of the thiostrepton resistance gene (tsr) of Streptomyces azureus: Transcriptional initiation from tsrp2 occurs after deletion of the — 35 region (1990)
    Springer
    Molecular genetics and genomics 221 (1990), S. 339-346 
    Details
    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
    URL: http://dx.doi.org/10.1007/BF00259397
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    Paper (German National Licenses)
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  • 3
    Electronic Resource
    Electronic Resource
    Glucose repression in Streptomyces coelicolor A3(2): a likely regulatory role for glucose kinase (1994)
    Springer
    Molecular genetics and genomics 244 (1994), S. 135-143 
    Details
    ISSN: 1617-4623
    Keywords: Streptomyces coelicolor A3(2) ; Glucose kinase ; Glucose repression ; Agarase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The glucose kinase gene (glkA-ORF3) of Streptomyces coelicolor A3(2) plays an essential role in glucose utilisation and in glucose repression of a variety of genes involved in the utilisation of alternative carbon sources. These genes include dagA, which encodes an extracellular agarase that permits agar utilisation. Suppressor mutants of glkA-ORF3 deletion strains capable of utilising glucose (Glc+) arise at a frequency of about 10−5 on prolonged incubation. The Glc+ phenotype of the mutants is reversible (at a frequency of about 10−3) and reflects either the activation of a normally silent glucose kinase gene or the modification of an existing sugar kinase. Although the level of glucose kinase activity in the Glc+ supressor mutants is similar to that in the glkA + parental strain, glucose repression of dagA remains defective. Expression of the glucose kinase gene of Zymomonas mobilis in glkA-ORF3 mutants restored glucose utilisation, but not glucose repression of dagA. Over-expression of glkA-ORF3 on a high-copy-number plasmid failed to restore glucose repression of dagA in glkA-ORF3 mutants and led to loss of glucose repression of dagA in a glkA + strain. These results suggest that glucose phosphorylation itself is not sufficient for glucose repression and that glkA-ORF3 plays a specific regulatory role in triggering glucose repression in S. coelicolor A3(2).
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00283514
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    Paper (German National Licenses)
    Fulltext
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