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18
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6844238927
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note
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+ gene.
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19
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6844263133
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note
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GST-Slp1 and GST were purified with the glutathione-Sepharose 4B beads (GSH) as recommended by the manufacturer (Pharmacia). An in vitro-translated Mad2 protein was obtained with the use of reticulocyte lysate (Promega).
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20
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6844262735
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note
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+ gene was inserted between two Sph I sites that were generated at ∼70 bp and 540 bp downstream of the putative translation initiation site of the gene.
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22
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6844231977
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note
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- leu1-32 ura4-d18) with ADH-M2. A detailed protocol will be available upon request.
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23
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6844225720
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note
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2+.
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24
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6844237869
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note
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A pair of oligonucleotides, GGGCATATGTTTGTAAATTCTATCAGCAGTGACGTT and GGGCATATGAGTGTTAAAGCGTCGTTTAGCCGGTGT, were used for PCR.
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26
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6844232810
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note
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GST and GST-Mad2 were expressed and the binding assay was carried out as described (16).
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27
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0026089183
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M. Glotzer, A. W. Murray, M. W. Kirschner, Nature 349, 132 (1991); A. Hershko, D. Ganoth, J. Pehrson, R. E. Palazzo, L. H. Cohen, J. Biol. Chem. 266, 4940 (1991); O. Cohen-Fix, J. Peters, M. W. Kirschner, D. Koshland, Genes Dev. 10, 3081 (1996); H. Funabiki et al., Nature 381, 438 (1996); Y.-L. Juang et al., Science 275, 1311 (1997).
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Zachariae, W.1
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S. Irniger, S. Piatti, C. Michaelis, K. Nasmyth, Cell 81, 269 (1995); R. W. King et al., ibid., p. 279; W. Zachariae, T. H. Shin, M. Galova, B. Obermaier, K. Nasmyth, Science 274, 1201 (1996); R. King, R. J. Deshaies, J. M. Peters, M. W. Kirschner, ibid., p. 1652.
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Science
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King, R.1
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Peters, J.M.3
Kirschner, M.W.4
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39
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6844222337
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note
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We thank G. Hannon and D. Beach for the cDNA library for the yeast two-hybrid system, P. Russell for pJL205, M. Yanagida for cut mutants and criticism of the manuscript, A. W. Murray for criticism of the manuscript and for sharing unpublished results, and G. Grills and D. Gebhard for technical assistance. D.P.L. was a Medical Student Research Fellow of the Howard Hughes Medical Institute and is supported by a training grant from the National Institutes of Health (5T32GM07491). A.K. was supported by the Human Frontier Science program. Supported by a grant from the National Institutes of Health (R01GM56305) to T.M.
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