Use of the Tn903 neomycin-resistance gene for promoter analysis in the fission yeast Schizosaccharomyces pombe |
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Authors: | Christine Lang-Hinrichs Claudia Dössereck Isabelle Fath Ulf Stahl |
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Institution: | (1) Institut für Gärungsgewerbe und Biotechnologie, Seestrasse 13, W-1000 Berlin 65, Germany;(2) FG Mikrobiologie und Genetik, Technische Universität Berlin, Gustav-Meyer-Allee 25, W-1000 Berlin 65, Germany |
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Abstract: | Summary The bacterial neo gene from transposon Tn903 (Tn601) was used for dominant transformation of the fission yeast Schizosaccharomyces pombe. It was found that high transformation efficiency was dependent on a high level of promoter activity, mediated by the strong promoter of the Schizosaccharomyces pombe alcohol dehydrogenase gene (adh1), as shown by comparing the efficiency of transformation to G418-resistance, the resistance levels of transformed cells, and the in vitro aminoglycoside phosphotransferase activity. On the other hand, the heterologous promoter of the Saccharomyces cerevisiae alcohol dehydrogenase I gene (adc1) is shown to be a weak promoter in Schizosaccharomyces pombe, though its activity is significantly enhanced in cells grown on glycerol as a carbon source. This system for selection and detection of promoter-active sequences may provide a useful basis for the analysis of promoter elements in fission yeast. |
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Keywords: | Schizosaccharomyces pombe S cerevisiae adc1 promoter S pombe adh1 promoter Aminoglycoside phosphotransferase |
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