title: Construction of Saccharomyces cerevisiae strain FAV20 useful in detection of immunosuppressants produced by soil actinomycetes.
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.
authors.affiliation: Institute of Molecular Genetics and Genetic Engineering, P.O. Box 23, Vojvode Stepe 444a, 11 001 Belgrade, Serbia and Montenegro.
authors.forename: Natasa
authors.initials: N
authors.lastname: Skoko
authors.forename: Jelena
authors.initials: J
authors.lastname: Vujovic
authors.forename: Miloje
authors.initials: M
authors.lastname: Savic
authors.forename: Natasa
authors.initials: N
authors.lastname: Papic
authors.forename: Branka
authors.initials: B
authors.lastname: Vasiljevic
authors.forename: Goran
authors.initials: G
authors.lastname: Ljubijankic
mesh_terms: D039903:Actinobacteria / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D004271:DNA, Fungal / Q000235:genetics; D007166:Immunosuppressive Agents / Q000378:metabolism / Q000494:pharmacology*; D008565:Membrane Proteins / Q000235:genetics; D008826:Microbial Sensitivity Tests; D016254:Mutagenesis, Insertional; D048630:Polyketide Synthases / Q000235:genetics; D016133:Polymerase Chain Reaction; D012441:Saccharomyces cerevisiae / Q000187:drug effects* / Q000235:genetics* / Q000254:growth & development; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics; D012988:Soil Microbiology; D016559:Tacrolimus / Q000378:metabolism / Q000494:pharmacology*; D014170:Transformation, Genetic
chemical_list: D004271:DNA, Fungal; D007166:Immunosuppressive Agents; D008565:Membrane Proteins; D029701:Saccharomyces cerevisiae Proteins; C095635:VMA22 protein, S cerevisiae; D048630:Polyketide Synthases; D016559:Tacrolimus