Dissection of Centromeric DNA from Yeast Yarrowia lipolytica and Identification of Protein-Binding Site Required for Plasmid Transmission(GENETICS, MOLECULAR BIOLOGY, AND GENE ENGINEERING)
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概要
- 論文の詳細を見る
In a dimorphic yeast, Yarrowia lipolytica, replicative plasmids can be established only in the coexistence of the replication origin(ORI) and centromere(CEN) from its own chromosomal DNA. Although six CEN sequences so far isolated from this yeast exhibit no similarity with conventional CEN DNA elements from other budding yeasts, they are confined within short regions(approximately 0.2 kb) and contain various conserved sequence blocks. We surveyed here a CENl1-1 sequence on an 0RI-containing plasmid by deletion and site-directed mutagenesis, and found a partial palindrome, CCTAATTTGG designated DS9, to be an essential element for high-efficiency transformation. In particular, point mutations that alter symmetry and/or length of the palindrome abrogated the activity of CEN1-1. Gel mobility shift assay of CEN1-1 DNA fragments incubated with Y. lipolytica nuclear proteins revealed four bands corresponding to protein-DNA complexes, whereas the mutations within DS9 that disabled transformation also abolished the formation of part of these complexes, depending on particular mutations. These results demonstrate that the palindrome is a binding site for specific protein(s) necessary for plasmid transmission in Y. lipolytica.
- 社団法人日本生物工学会の論文
- 2008-06-25
著者
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NAGAHAMA KAZUHIRO
Department of Applied Microbial Technology, Faculty of Biotechnology and Life Science, Sojo Universi
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MATSUOKA MASAYOSHI
Department of Applied Microbial Technology, Faculty of Biotechnology and Life Science, Sojo Universi
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OGAWA TAKAHIRA
Department of Applied Microbial Technology, Faculty of Biotechnology and Life Science, Sojo Universi
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Sakai Hiroaki
Department Of Thoracic Surgery Kyoto University Hospital
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中村 薫
京大 化研
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中村 厚三
Univ. Tokyo Tokyo Jpn
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Ogawa T
Shinshu Univ. Nagano Jpn
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Ogawa T
Kyoto Univ. Kyoto Jpn
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Ogawa Takahiro
Department Of Biological Science And Technology Science University Of Tokyo
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Ogawa T
Department Of Applied Microbial Technology Faculty Of Biotechnology And Life Science Sojo University
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Matsuoka M
Department Of Applied Microbial Technology Faculty Of Biotechnology And Life Science Sojo University
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Matsuoka Masayoshi
Dept. Applied Microb. Technol. Sojo Univ.
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Nagahama Kazuhiro
Department Of Applied Microbial Technology Faculty Of Biotechnology And Life Science Sojo University
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Yamane Takayoshi
Department of Applied Microbial Technology, Faculty of Biotechnology and Life Science, Sojo Universi
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Sakai Hiroaki
Department Of Applied Microbial Technology Faculty Of Biotechnology And Life Science Sojo University
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Ogawa Tomoya
Laboratory Of Synthetic Cellulary Chemistry The Institute Of Physical And Chemical Research (riken)
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Ogawa Takahira
Department Of Applied Microbial Technology Faculty Of Biotechnology And Life Science Sojo University
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Ogawa Takahira
Dept. Appl. Microbial Technology Kumamoto Institute Of Technology
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Nagahama Kazuhiro
Department Of Applied Microbial Technology The Kumanoto Institute Of Technology
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Matsuoka Masayoshi
Dept. Appl. Microbial Technology Kumamoto Institute Of Technology
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Matsuoka Masayoshi
Department Of Applied Microbial Technology Faculty Of Biotechnology And Life Science Sojo University
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Sakai Hiroaki
Department Of Anesthesiology Juntendo University Shizuoka Hospital
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Ogawa Tadashi
Graduate School Of Bioagricultural Sciences Nagoya University
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Okayama T
Research Institute Daiichi Fine Chemical Co. Ltd.
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Nakamura Kahoru
Department Of Nutrition Tokyo University Of Agriculture
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Yamane Takayoshi
Department Of Applied Microbial Technology Faculty Of Biotechnology And Life Science Sojo University
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