Mycobiology.  2003 Mar;31(1):42-45. 10.4489/MYCO.2003.31.1.042.

Transformation of the Edible Basidiomycete, Pleurotus ostreatus to Phleomycin Resistance

Affiliations
  • 1Molecular Physiology Division, National Institute of Agricultural Biotechnology, Suweon 441-707 Korea. bgkimyes@rda.go.kr
  • 2Department of Biochemistry, Kon-Kuk University, Chung-ju 380-701, South Korea.
  • 3Applied Microbiology Division, National Institute of Agricultural Science and Trechnology, Suweon 441-707 Korea.
  • 4Forestry and Forest Product Research Institute, P.O. Box 16, Ibaraki, 305-8687 Japan.

Abstract

For transformation of Pleurotus ostreatus, two novel vectors, pPhKM1 and pPhKM2, were constructed, using the regulatory sequences of the P. sajor-caju beta-tubulin gene (TUB1) and the ble gene encoding phleomycin binding protein. pPhKM1 contains ble fused to the TUB1 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 pPhKM1 or pPhKM2. This transformation system generated stable phleomycin-resistant transformants.

Keyword

Basidiomycete; Phleomycin resistance; Pleurotus ostreatus; Transformation; beta-tublin

MeSH Terms

Basidiomycota*
Carrier Proteins
DNA
Phleomycins*
Pleurotus*
Schizophyllum
Terminator Regions, Genetic
Tubulin
Carrier Proteins
DNA
Phleomycins
Tubulin
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