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[Primers for (1,3)-β-glucan synthase gene amplification and partial characterization of the enzyme in Ganoderma
Jessica Valeria Guerrero-Torres1, Gerardo Mata, Daniel Martínez-Carrera
1Departamento de Bioingeniería, Unidad Profesional Interdisciplinaria de Biotecnología, Instituto Politécnico Nacional, México, D. F., México.
Background:
β-(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.
Aims:
To 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.
Methods:
The 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.
Results:
Three 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.
Conclusions:
The 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.
