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タイトル: | Absence of a gene encoding cytosine deaminase in the genome of the agaricomycete Coprinopsis cinerea enables simple marker recycling through 5-fluorocytosine counterselection. |
著者: | Nakazawa, Takehito https://orcid.org/0000-0002-9654-2124 (unconfirmed) Honda, Yoichi https://orcid.org/0000-0002-7218-7367 (unconfirmed) |
著者名の別形: | 中沢, 威人 |
キーワード: | basidiomycete Coprinopsis cinerea Pleurotus ostreatus FCY1 pyrG cytosine deaminase |
発行日: | Aug-2015 |
出版者: | Oxford University Press |
誌名: | FEMS microbiology letters |
巻: | 362 |
号: | 15 |
論文番号: | fnv123 |
抄録: | Coprinopsis cinerea is a model species for molecular genetics studies of sexual development in agaricomycetes or homobasidiomycetes. Recently, efficient gene targeting was established in this fungus by generating Cc.ku70 or Cc.lig4 disruptants. To determine the molecular mechanisms underlying sexual development, which involves many genes, generating multiple gene disruptants is required. However, the number of transformation markers available for C. cinerea is limited. This problem would be solved by establishing marker recycling. In this study, we found that C. cinerea lacks a gene encoding a homolog of Saccharomyces cerevisiae cytosine deaminase (Fcy1p) in its genome, which is present in many other fungi. We also observed that C. cinerea is resistant to 5-fluorocytosine. Based on these findings, we established a simple marker recycling method in this fungus using 5-fluorocytosine counterselection after heterologous expression of FCY1 derived from Pleurotus ostreatus, together with the hygromycin resistance gene. This study proposes a simple genetic manipulation system that can be performed using wild-type strains of several fungi that lack a gene homologous to S. cerevisiae FCY1 in their genomes. |
記述: | First published online: 27 July 2015 |
著作権等: | This is a pre-copyedited, author-produced PDF of an article accepted for publication in 'FEMS Microbiology Letters' following peer review. The version of record [Takehito Nakazawa, Yoichi Honda. Absence of a gene encoding cytosine deaminase in the genome of the agaricomycete Coprinopsis cinerea enables simple marker recycling through 5-fluorocytosine counterselection. 362. 15. fnv123] is available online at: http://femsle.oxfordjournals.org/content/362/15/fnv123. The full-text file will be made open to the public on 27 July 2016 in accordance with publisher's 'Terms and Conditions for Self-Archiving'. この論文は出版社版でありません。引用の際には出版社版をご確認ご利用ください。 This is not the published version. Please cite only the published version. |
URI: | http://hdl.handle.net/2433/207354 |
DOI(出版社版): | 10.1093/femsle/fnv123 |
PubMed ID: | 26223587 |
出現コレクション: | 学術雑誌掲載論文等 |
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