Use of the Tn903 neomycin-resistance gene for promoter analysis in the fission yeast Schizosaccharomyces pombe.
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 amino-glycoside 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.
Lang-Hinrichs
C
C
Institut für Gärungsgewerbe und Biotechnologie, Berlin, Federal Republic of Germany.
Dössereck
C
C
Fath
I
I
Stahl
U
U
D000426:Alcohol Dehydrogenase / Q000235:genetics / Q000378:metabolism; D015152:Blotting, Northern; D015139:Blotting, Southern; D003001:Cloning, Molecular; D004251:DNA Transposable Elements; D004352:Drug Resistance, Microbial / Q000235:genetics; D015966:Gene Expression Regulation, Fungal; D005798:Genes, Bacterial; D019868:Kanamycin Kinase; D009355:Neomycin / Q000494:pharmacology*; D010770:Phosphotransferases / Q000235:genetics / Q000378:metabolism; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics; D012568:Schizosaccharomyces / Q000201:enzymology / Q000235:genetics*; D014170:Transformation, Genetic
D004251:DNA Transposable Elements; D000426:Alcohol Dehydrogenase; D010770:Phosphotransferases; D019868:Kanamycin Kinase; D009355:Neomycin