Tn5099, a xylE promoter probe transposon for Streptomyces spp

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Construction and application of plasmid- and transposon-based promoter-probe vectors for Streptomyces spp. that employ a Vibrio harveyi luciferase reporter cassette.

Several versatile promoter-probe vectors have been constructed for Streptomyces strains which utilize the production of blue-green light as a measure of transcription activity. Three plasmid vectors (two high and one low copy number) and two transposons are described. The multicopy plasmids pRS1106 and pRS1108 contain a transcription terminator and multiple-cloning polylinker upstream of promot...

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TnphoA: a transposon probe for protein export signals.

We constructed a derivative of transposon Tn5 that permits the generation of hybrid proteins composed of alkaline phosphatase (EC 3.1.3.1) lacking its signal peptide fused to amino-terminal sequences of other proteins. Such a hybrid gives alkaline phosphatase activity if the protein fused to alkaline phosphatase contributes sequences that promote export and thus compensate for the missing alkal...

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Transposon mutagenesis of marine Vibrio spp.

Coliphage P1 was used to transduce derivatives of transposons Tn5 and mini-Mu into marine Vibrio spp. Transposon Tn5 encoding tetracycline resistance (Tn5-132) was used to isolate mutants of Vibrio harveyi defective in genes for bioluminescence (lux). Insertion of transposon Tn5-132 into the lux gene region was demonstrated by intraspecific transduction with phage hv-1 and by Southern blot hybr...

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Development of a temperature-inducible expression system for Streptomyces spp.

PCR mutagenesis of a 0.9-kbp fragment, containing a repressor gene, traR, and its target promoter, Ptra, from Streptomyces nigrifaciens plasmid pSN22, produced Streptomyces lividans clones with temperature-inducible Ptra expression. Using the promoterless gene for the thermostable Thermus flavus malate dehydrogenase as an indicator, an induction of enzyme activity of as much as was observed in ...

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Nutrient effects on ten Streptomyces spp. sporulation

The effects of nutrients on sporulation of ten Streptomyces spp. was studied on 26 media. The media included starch-casein-KNO3, glycerol-arginine (GA), actinomycete-isolation (AI), tryptone-yeast extractglucose (TYG), and tryptone-soy agars. Media compositions had complicated effects on sporulation. The sporulation on media with minerals of GA agar and yeast extract was usually better than on ...

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ژورنال

عنوان ژورنال: Journal of Bacteriology

سال: 1991

ISSN: 0021-9193,1098-5530

DOI: 10.1128/jb.173.17.5573-5577.1991