Transformation of the Edible Basidiomycete, Pleurotus ostreatus to Phleomycin Resistance

Abstract

For transformation of Pleurotus ostreatus, two novel vectors, pPhKM1 and pPhKM2, were constructed, using the regulatory sequences of the P. sajor-caju β-tubulin gene (TUBI) and the ble gene encoding phleomycin binding protein. pPhKM1 contains ble fused to the TUBI promoter and the Schizophyllum commune GPD terminator. pPhKM2 contains ble fused to the promoter and terminator regions of P. sajor-caju TUB1. To confirm phleomycin-resistance activity, each vector was cotransformed with pTRura3-2 into the P. ostreatus homokaryotic ura⁻ strain. The transforming DNA was stably integrated into the genomic DNA. Subsequently, phleomycin resistance was conferred on wild-type dikaryotic P. ostreatus by transformation with pPhKMl or pPhKM2. This transformation system generated stable phleomycin-resistant transformants.

Keywords::

• Basidiomycete
• Phleomycin resistance
• Pleurotus ostreatus
• Transformation
• β-tublin