Construction of Saccharomyces cerevisiae strain FAV20 useful in detection of immunosuppressants produced by soil actinomycetes |
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Authors: | Skoko Natasa Vujovic Jelena Savic Miloje Papic Natasa Vasiljevic Branka Ljubijankic Goran |
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Affiliation: | Institute of Molecular Genetics and Genetic Engineering, P.O. Box 23, Vojvode Stepe 444a, 11 001 Belgrade, Serbia and Montenegro. |
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Abstract: | The screening of microbial natural products continues to represent an important route to the discovery of novel chemicals for development of new therapeutic agents. The aim of this work was to develop an efficient method for the detection of immunosuppressive compounds produced by soil actinomycetes. Mutant strain of Saccharomyces cerevisiae, named FAV20, sensitive to FK506 was constructed by disrupting VMA22 gene using the selectable marker kanMX4 which allowed detection of integration events. Actinomycetes were isolated from different soil samples and in a newly developed test with S. cerevisiae FAV20, six strains have been identified that produce bioactive compounds with the same mechanism of action as FK506. S. cerevisiae FAV20 can be easily used as a test strain in drug screening programs based on inhibition of the calcineurin phosphatase dependent signaling pathway in the cell. |
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