Transformation of the Edible Basidiomycete Lentinus edodes by Restriction Enzyme-Mediated Integration of Plasmid DNA

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概要

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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 2.5μg of DNA. The conventional PEG transformation without SalI, however, yielded only 1.5 transformants per 25μg 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.

• 社団法人日本農芸化学会の論文
• 1998-12-23

著者

• Sato Takahide

  Department Of Bioproduction Science Faculty Of Horticulture Chiba University

• Hirano T

  Kyoto Univ. Kyoto Jpn

• Enei H

  Iwate Biotechnol. Res. Center Iwate Jpn

• Enei Hitoshi

  Iwate Biotechnology Research Center

• Sato Takako

  The Deepstar Group Japan Marine Science And Technology Center

• Yaegashi Kaori

  Iwate Biotechnology Res. Center

• Sato T

  Institute Of Applied Biochemistry University Of Tsukuba

• SHISHIDO Kazuo

  Department of Life Science, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Tech

• KAJIWARA Susumu

  Department of Life Science, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Tech

• Enei H

  Iwate Biotechnology Research Center

• SATO Toshitsugu

  Iwate Biotechnology Research Center

• HIRANO Tatsuya

  Iwate Biotechnology Research Center

• ISHII Shizuko

  Iwate Biotechnology Research Center

• Ishii S

  San-ei Sucrochemical Co. Ltd. Chiba Jpn

• Shishido Kazuo

  Department Of Life Science Faculty Of Bioscience And Biotechnology Tokyo Institute Of Technology

• Sato T

  Iwate Biotechnology Research Center:department Of Biotechnology And Environmental Chemistry National

• Kajiwara Susumu

  Department Of Life Science Faculty Of Bioscience And Biotechnology Tokyo Institute Of Technology

• Taniyama Hiroko

  Department Of System Engineering Of Materials And Life Science Faculty Of Engineering Toyama Univers

• Horikawa Takashi

  Collaborative Research Center For Advanced Science And Technology Osaka University

• Sato Tsutomu

  Graduate School Of Science And Technology Faculty Of Agriculture Niigata University

• Sato Toshitugu

  Iwate Biotechnology Research Center

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