Departamento de Ingeniería Genética, Unidad Irapuato, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional, Irapuato, Guanajuato, Mexico.
FEMS Microbiol Lett. 2010 Nov;312(1):77-83. doi: 10.1111/j.1574-6968.2010.02099.x. Epub 2010 Sep 15.
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.
通过计算机分析,我们证明了一个之前描述的嵌合基因(Spe-Sdh),它编码精胺合酶,这是一种参与多胺合成的关键酶,和 saccharopine 脱氢酶,一种参与真菌中赖氨酸合成的酶,仅存在于所有担子菌亚门的成员中,但不存在于任何其他生物群体中。我们利用这一特点设计了简并引物,通过 PCR 扩增 Spe-Sdh 基因的特定片段,作为一种工具来明确鉴定担子菌分离物。该程序的特异性使用不同的真菌物种进行了测试。正如预期的那样,只有担子菌物种得到了阳性结果,而其他真菌门的物种则没有扩增。