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Molecular Breeding

, Volume 1, Issue 1, pp 79–89 | Cite as

Construction and characterization of a rice YAC library for physical mapping

  • Yosuke Umehara
  • Akiko Inagaki
  • Hiroshi Tanoue
  • Yuji Yasukochi
  • Yoshiaki Nagamura
  • Shoko Saji
  • Yoshiaki Otsuki
  • Tatsuhito Fujimura
  • Nori Kurata
  • Yuzo Minobe
Article

Abstract

Genomic libraries of rice,Oryza sativa L. cv. Nipponbare, in yeast artificial chromosomes were prepared for construction of a rice physical map. High-molecular-weight genomic DNA was extracted from cultured suspension cells embedded in agarose plugs. After size fractionation of theEco RI- andNot I-digested DNA fragments, they were ligated with pYAC4 and pYAC55, respectively, and used to transformSaccharomyces cerevisiae AB1380. A total of 6932 clones were obtained containing on average ca. 350 kb DNA. The YAC library was estimated to contain six haploid genome equivalents. The YACs were examined for their chimerism by mapping both ends on an RFLP linkage map. Most YACs withEco RI fragments below 400 kb were intact colinear clones. About 40% of clones were chimeric. Genetic mapping of end clones from large size YACs revealed that the physical distance corresponding to 1 cM genetic distance varies from 120 to 1000 kb, depending on the chromosome region. To select and order YAC clones for making contig maps, high-density colony hybridization using ECL was applied. With several probes, at least one and at most ten YAC clones could be selected in this library. The library size and clone insert size indicate that this YAC library is suitable for physical map construction and map-based cloning.

Key words

physical mapping rice YAC 

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Copyright information

© Kluwer Academic Publishers 1995

Authors and Affiliations

  • Yosuke Umehara
    • 1
  • Akiko Inagaki
    • 1
  • Hiroshi Tanoue
    • 1
  • Yuji Yasukochi
    • 1
  • Yoshiaki Nagamura
    • 1
  • Shoko Saji
    • 1
  • Yoshiaki Otsuki
    • 2
  • Tatsuhito Fujimura
    • 1
  • Nori Kurata
    • 1
  • Yuzo Minobe
    • 1
  1. 1.Rice Genome Research ProgramNational Institute of Agrobiological Resources/Institute of Society for Techno-innovation of Agriculture, Forestry and FisheriesTsukuba, IbarakiJapan
  2. 2.National Agricultural Research CenterTsukuba, IbarakiJapan

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