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http://dx.doi.org/10.4489/MYCO.2003.31.1.032

The Efficient Transformation of Pleurotus ostreatus using REMI Method  

Joh, Joong-Ho (Department of Biochemistry, Kon-kuk University)
Kim, Beom-Gi (Molecular physiology division, National Institute of Agricultural Biotechnology)
Chu, Kyo-Sun (Department of Biochemistry, Kon-kuk University)
Kong, Won-Sik (Applied microbiology division, National Institute of Agricultural Sciences and Technologies)
Yoo, Young-Bok (Applied microbiology division, National Institute of Agricultural Sciences and Technologies)
Lee, Chang-Soo (Department of Biochemistry, Kon-kuk University)
Publication Information
Mycobiology / v.31, no.1, 2003 , pp. 32-35 More about this Journal
Abstract
Restriction enzyme-mediated integration(REMI) was used to transform uracil auxotrophs of Pleurotus ostreatus to prototrophy. When protoplasts of Pleurotus ostreatus were treated by the reaction mixture containing 10 units of BamHI, the frequency of REMI was about 64 transformants per 1 ${\mu}g$ of DNA. This efficiency was increased by 14.2 times compared with that of the conventional PEG transformation. The optimal condition for REMI of P. ostreatus was achieved when 1 ${\mu}g$ of linearized pTRura3-2 DNA was added into $1{\times}10^7$ protoplasts along with 10 units BamHI. Southern blot analysis revealed that about 50% of transformants examined were caused by REMI event and 30% carried single copy insertion at the genome. This suggested that the REMI method might be a useful tool for efficient transformation and tagging mutagenesis of P. ostreatus.
Keywords
Pleurotus ostreatus; Restriction enzyme-mediated integration(REMI); Uracil auxotrophic marker;
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1 Berges, T and Barreau, C. 1991. Isolation of uridine auxotrophs from Trichoderma reesei and efficient transformation with the cloned ura3 and ura5 genes. Curro Genet. 19: 359-365   DOI   ScienceOn
2 Bolker, M., Bohnert, H. U., Braun, K. H., Gorl, J. and Kahmann, R. 1995. Tagging pathogenicity genes in Ustilago maydis by restriction enzyme-mediated integration (REMI). Mol. Gen. Genet. 248: 547-552   DOI   ScienceOn
3 Brown, D. H. Jr., Slobodkin, I. V. and Kumamoto, C. A. 1996. Stable transformation and regulated expression of an inducible reporter construct in Candida albicans restriction enzymemediated integration. Mol. Gen. Genet. 251: 75-80
4 Cummings, W. J., Celerin, M., Crodian, J., Brunick, L. K and Zolan, M. E. 1999. Insertional mutagenesis in Coprinus cinereus: use of a dominant selectable marker to generate tagged, sporulation-defective mutants, Curro Genet. 36(6): 371-38
5 Granado, J. D., Kertesz-Chaloupkova, K, Aebi, M. and Kues, D. 1997. Restriction enzyme-mediated DNA integration in Coprinus cinereus. Mol. Gen. Genet. 256: 28-36   DOI   ScienceOn
6 Honda, Y, Matsuyama, T, Irie, T, Watanabe, T and Kuwahara, M. 2000. Carboxin resistence transformation of the hornobasidiomycete fungus Pleurotus ostreatus. Curro Genet. 37: 209-212   DOI   ScienceOn
7 Irie, T, Honda, Y, Watanabe, T. and Kuwahara, M. 2001. Efficient transformation of filamentous fungus Pleurotus ostreatus using single-strand carrier DNA. Appl. Microbiol. Biotechnol. 55: 563-565   DOI   ScienceOn
8 Jia, J. H., Buswell-John, A. and Peberdy, J. F. A. 1998. Transformation of the edible fungi, Pleurotus ostreatus and Volvariella Volvacea. Mycol. Res. 102: 876-880   DOI   ScienceOn
9 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. FEMS Micobiol. Lett. 181: 225-228   DOI   ScienceOn
10 Kuspa, A. and Loomis, W. F. 1992. Tagging developmental genes in Dictyostelium by restriction enzyme-mediated integration of plasmid DNA. Proc. Natl. Acad. Sci. USA 89: 8803-8807   DOI   ScienceOn
11 Lu, S., Lyngholm, L., Yang, G., Bronson, C., Yoder, O. C. and Turgeon, B. G. 1994. Tagged mutations at the ToxI locus of Cochliobolus heterostrophus by restriction enzyme-mediated integration. Proc. Natl. Acad. Sci. USA 91: 12649-12653   DOI   ScienceOn
12 Noel, T. and Labarere, J. 1994. Homologous transformation of the edible basidiomycete Agrocybe aegerita, with the URA1 gene: Characterization of integrative events and of rearranged free plasmid in transfonnants. Curro Genet. 25: 432-437   DOI   ScienceOn
13 Peng, M., Lemke, P. A. and Shaw, J. J. 1993. Improved conditions for protoplast formation and transformation of Pleurotus ostreatus. Appl. Microbiol. Biotechnol. 40: 101-106
14 Reader, D. and Broda, P. 1985. Rapid preparation of DNA from filamentous fungi. Lett. Appl. Microbiol. 1: 17-20   DOI
15 Rhee, M. D. van de, Graca, P. M. A., Huizing, H. and Mooibrock, H. 1996. Transformation of the cultivated mushroom, Agaricus bisporus, to hygromycin B resistance. Mol. Gen. Genet. 250: 252-258
16 Sanchez, O., Navarro, R. E. and Aguirre, J. 1998. Increased transformation frequency and tagging of developmental genes in Aspergillus nidulans by restriction enzyme-mediated integration. Mol. Gen. Genet. 258: 89-94   DOI   ScienceOn
17 Vyas, B. R. M., Bakowski, S., Sasek, V. and Matucha, M. 1994. Degradation of anthracene by selected white rot fungi. FEMS Microbiol. Ecol. 14: 65-70   DOI   ScienceOn
18 Sato, T, Yaegashi, K, Ishii, S., Hirano, T, Kajiwara, S., Shishido, K and Enei, H. 1998. Transformation of the edible basidiomycete Lentinus edodes by restriction enzyme-mediated integration of plasmid DNA. Biosci. Biotechnol. Biochem. 62: 2346-2350   DOI   ScienceOn
19 Schiestl, R. H. and Petes, T D. 1991. Integration of DNA fragments by illegitimate recombination in Saccharomyces cerevisiae. Proc. Natl. Acad. Sci. USA 88: 7585-7589   DOI   ScienceOn
20 Shi, Z., Christian, D. and Leung, H. 1995. Enhanced transformation in Magnapothe grisea by restriction enzyme mediated integration of plasmid DNA. Phytopathology 85: 329-333   DOI   ScienceOn
21 Yanai, K, Yonekura, K, Usami, H., Hirayama, M., Kajiwara, S., Yamazaki, T, Shishido, K and Adachi, T. 1996. The integrative transformation of Pleurotus ostreatus using bialaphos resistance as a dominant selectable marker. Biosci. Biotechnol. Biochem. 60: 472-475   DOI   ScienceOn
22 Akamatsu, H., Itoh, Y, Kodama, M., Otani, H. and Kohmoto, K 1997. AAL-toxin deficient mutants of A. alternata tomato pathotype by restriction enzyme mediated integration. Phytopathology 87: 967-972