caa a22 4500 46575788X CHVBK 20180323111852.0 cr unu---uuuuu 170327e19901101xx s 000 0 eng 10.1007/BF00318213 doi (NATIONALLICENCE)springer-10.1007/BF00318213 Construction and characterization of centric circular and acentric linear chromosomes in fission yeast [Elektronische Daten] [Tomohiro Matsumoto, Shin Murakami, Osami Niwa, Mitsuhiro Yanagida] Summary: Gene disruption and gap repair of chromosomal DNA have been frequently employed techniques in yeast genetics. To extend the possibility of using these gene manipulations for larger genomic regions, we have examined the maximal sizes of chromosomal DNA disrupted or repaired in vivo. Here we report a simple, potentially general, method for selectively deleting a 150 kb region, or gap-filling a 100 kb region, in the fission yeast genome. This enables the generation of acentric linear chromosomes by deletion, or the cloning of large functional centromeric DNAs into circular minichromosomes by gap-filling. The fidelity of the resulting gap-filling is high, judging from partial-digestion mapping of gap-repaired DNAs. By analysing a series of such circular minichromosomes, we conclude that only a part of the repetitive centromeric region, including the central domain, is essential for mitotic and meiotic chromosome segregation. Acentric linear chromosomes, although unstable, could be maintained, indicating that it may be possible to construct an acentric vector for large DNA fragments in this organism. Springer-Verlag, 1990 Artificial chromosome nationallicence Centromere nationallicence Genome manipulation nationallicence Schizosaccharomyces pombe nationallicence Matsumoto Tomohiro Department of Biophysics, Faculty of Science, Kyoto University, Kitashirakawa, Sakyo-ku, 606, Kyoto, Japan aut Murakami Shin Department of Biophysics, Faculty of Science, Kyoto University, Kitashirakawa, Sakyo-ku, 606, Kyoto, Japan aut Niwa Osami Department of Biophysics, Faculty of Science, Kyoto University, Kitashirakawa, Sakyo-ku, 606, Kyoto, Japan aut Yanagida Mitsuhiro Department of Biophysics, Faculty of Science, Kyoto University, Kitashirakawa, Sakyo-ku, 606, Kyoto, Japan aut Current Genetics Springer-Verlag 18/4(1990-11-01), 323-330 0172-8083 18:4<323 1990 18 294 https://doi.org/10.1007/BF00318213 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF00318213 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Matsumoto Tomohiro Department of Biophysics, Faculty of Science, Kyoto University, Kitashirakawa, Sakyo-ku, 606, Kyoto, Japan aut NATIONALLICENCE 700 1- Murakami Shin Department of Biophysics, Faculty of Science, Kyoto University, Kitashirakawa, Sakyo-ku, 606, Kyoto, Japan aut NATIONALLICENCE 700 1- Niwa Osami Department of Biophysics, Faculty of Science, Kyoto University, Kitashirakawa, Sakyo-ku, 606, Kyoto, Japan aut NATIONALLICENCE 700 1- Yanagida Mitsuhiro Department of Biophysics, Faculty of Science, Kyoto University, Kitashirakawa, Sakyo-ku, 606, Kyoto, Japan aut NATIONALLICENCE 773 0- Current Genetics Springer-Verlag 18/4(1990-11-01), 323-330 0172-8083 18:4<323 1990 18 294 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer