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| | ONLINE | ISSN | : | 1347-6947 | | PRINT | ISSN | : | 0916-8451 |
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| | Bioscience, Biotechnology, and Biochemistry |
| Vol. 62 (1998) , No. 12 pp.2346-2350 |
| [Image PDF (1896K)] [References]  |
 | Transformation of the Edible Basidiomycete Lentinus edodes by Restriction Enzyme-Mediated Integration of Plasmid DNA
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| | 1) Iwate Biotechnology Research Center
2) Department of Life Science, Faculty of Bioscience and Biotechnology, Tokyo Institute of Technology |
| (Received July 2, 1998)
(Accepted August 26, 1998)
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| | We have used the restriction enzyme-mediated DNA integration (REMI) method to establish a transformation system in Lentinus edodes using the recombinant plasmid pLC1-hph, which contains the L. edodes transcriptional signals and an Escherichia coli hygromycin B phosphotransferase gene. Protoplasts of L. edodes were treated by the PEG transformation mixture containing 50 units of SalI, which cleaves pLC1-hph at a single site, yielding about 15 transformants per 2.5 μg of DNA. The conventional PEG transformation without SalI, however, yielded only 1.5 transformants per 25 μg of DNA. The optimal amount of SalI for increased transformation was 50 units. In the case of transformation with SphI, which cleaves the plasmid at one site, the optimal amount of the enzyme was 2.5 units. Southern blot analysis of the SphI-derived transformants suggested that 50% of the plasmid integrations were REMI events.
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| To cite this article: | | Toshitsugu SATO, Kaori YAEGASHI, Shizuko ISHII, Tatsuya HIRANO, Susumu KAJIWARA, Kazuo SHISHIDO and Hitoshi ENEI, “Transformation of the Edible Basidiomycete Lentinus edodes by Restriction Enzyme-Mediated Integration of Plasmid DNA”, Biosci. Biotechnol. Biochem., Vol. 62, 2346-2350 (1998) . | |
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| doi:10.1271/bbb.62.2346 | | JOI JST.JSTAGE/bbb/62.2346 |
| Copyright (c) 2005 by Japan Society for Bioscience, Biotechnology, and Agrochemistry |
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