Cebadores para la amplificación del gen de la (1,3)-β-glucano sintasa y caracterización parcial de la enzima en Ganoderma lucidum

Abstract

Backgroundβ-(1,3)(1,6)-D-glucan is fungal cell wall component that has demonstrated immunomodulatory and anti-cancer effects. The (1,3)-β-glucan synthase is one of the main enzymes involved in its biosynthesis. AimsTo design primers to partially amplify and characterize the (1,3)-β-glucan synthase gene and to determine them in Ganoderma lucidum (G. Lucidum) strain CP-132. MethodsThe primers were designed on the basis of homologous genes in other fungi. Then, using the PCR technique, primers were tested using DNA extracted from the G. lucidum strain CP-382. Amplified sequences were compared with those from the GenBank. ResultsThree primer pairs were designed; all of them produced amplicons of the expected size. The sequences obtained with primer pairs BGS2113UmF and BGS3097UmR, and BGS547UmF and BGS2113UmR matched with 2 sections of the (1,3)-β-glucan synthase gene. The deduced amino acid sequences showed high similarity with homologous genes from other fungi, particularly with those of the Agaricomycetes class. ConclusionsThe primer design to partially amplify the (1,3)-β-glucan synthase gene of G. lucidum using sequences from homologous genes was successful. These primers will allow to characterize this important enzyme in a wide group of fungi.