A molecular probe for Basidiomycota: the spermidine synthase-saccharopine dehydrogenase chimeric gene.
By means of an in silico analysis, we demonstrated that a previously described chimeric gene (Spe-Sdh) encoding spermidine synthase, a key enzyme involved in the synthesis of polyamines, and saccharopine dehydrogenase, an enzyme involved in lysine synthesis in fungi, were present exclusively in members of all Basidiomycota subphyla, but not in any other group of living organisms. We used this feature to design degenerated primers to amplify a specific fragment of the Spe-Sdh gene by PCR, as a tool to unequivocally identify Basidiomycota isolates. The specificity of this procedure was tested using different fungal species. As expected, positive results were obtained only with Basidiomycota species, whereas no amplification was achieved with species belonging to other fungal phyla.
León-Ramírez
Claudia G
CG
Departamento de Ingeniería Genética, Unidad Irapuato, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional, Irapuato, Guanajuato, Mexico.
Valdés-Santiago
Laura
L
Campos-Góngora
Eduardo
E
Ortiz-Castellanos
Lucila
L
Aréchiga-Carvajal
Elva T
ET
Ruiz-Herrera
José
J
D000595:Amino Acid Sequence; D001483:Base Sequence; D001487:Basidiomycota / Q000737:chemistry / Q000145:classification / Q000201:enzymology* / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D015335:Molecular Probes / Q000235:genetics*; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D012443:Saccharopine Dehydrogenases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D016415:Sequence Alignment; D013045:Species Specificity; D011436:Spermidine Synthase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
D017931:DNA Primers; D005656:Fungal Proteins; D015335:Molecular Probes; D012443:Saccharopine Dehydrogenases; D011436:Spermidine Synthase