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http://dx.doi.org/10.4489/MYCO.2003.31.1.042

Transformation of the Edible Basidiomycete, Pleurotus ostreatus to Phleomycin Resistance  

Kim, Beom-Gi (Molecular Physiology Division, National Institute of Agricultural Biotechnology)
Joh, Jung-Ho (Department of Biochemistry, Kon-Kuk University)
Yoo, Young-Bok (Applied Microbiology Division, National Institute of Agricultural Science and Trechnology)
Magae, Yumi (Forestry and Forest Product Research Institute)
Publication Information
Mycobiology / v.31, no.1, 2003 , pp. 42-45 More about this Journal
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 cotrans-formed 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.
Keywords
Basidiomycete; Phleomycin resistance; Pleurotus ostreatus; Transformation; $\beta$-tublin;
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1 Bartholomew, K., Santos, G. D., Dumonceaux, T., Charles, T and Archibald, F. 2001. Genetic transformation of Trametes versicolor to phleomycin resistance with the dominant selectable marker shble. Appl. Microbiol. Biotechnol. 56: 201-204   DOI   ScienceOn
2 Berges, T and Barreau, C. 1991. Isolation of uridine auxotrophs from Trichoderma reesei and efficient transformation with thecloned ura3 and ura5 genes. Curro Genet. 19: 359-365   DOI   ScienceOn
3 Chang, S.-T and Miles, P. G. 1989. Edible mushrooms and their cultivation CRC Press Inc, pp. 165-166
4 Honda, Y., Matsuyama, T, Irie, T, Watanabe, T and Kuwahara, M. 2000 Carboxin resistance transformation of the homobasidiomycete fungus Pleurotus ostreatus. Curro Genet. 37: 209-212
5 Irie, T, Honda, Y, Hirano, T, Sato, T, Enei, H., Watanabe, T and Kuwahara, M. 2001. Stable transformation of Pleurotus ostreatus to hygromycin B resistance using Lentinus edodes GPD expression signals. Appl. Microbiol. Biotechnol. 56: 707-709   DOI   ScienceOn
6 Kim, B.-G., Magae, Y, Yoo, Y-B. and Kwon, S.-T 1999. Isolation and transformation of uracil auxotrophs of the edible basidiomycete Pleurotus ostreatus. EFMS Microbial. Lett. 181: 225-228   DOI   ScienceOn
7 Peng, M., Lemke, P. A. and Singh, N. K. 1993. A nucleotide sequence involved in replicative transformation of a filamentous fungus. Curro Genet. 24: 114-121   DOI   ScienceOn
8 Raeder, U. and Broda, P. 1985. Rapid preparation of DNA from filamentous fungi. Lett. Appl. Microbiol. 1: 17-20   DOI
9 Schuren, F. H. and Wessels, J. G. 1994. Highly-efficient transformation of the homobasidiomycete Schirophyllum commune to phleomycin resistance. Curro Genet. 26: 179-183   DOI   ScienceOn
10 Stappert, J. 1994. Methods for generating multiple site-directed mutations in vitro. In: Griffin, H. G., Griffin, A. M. PCR Technology Current Innovations. eRC Press Inc, pp. 59-65
11 Kim, B.-G., Yoo, Y-B., Kwon, S.-T and Magae, Y. 2001. Molecular characterization of -tubulin gene from Pleurotus sajor-caju. Biosci. Biotechnol. Biochem. 65: 2280-2283   DOI   ScienceOn