Theoretical and Applied Genetics

, Volume 94, Issue 2, pp 170–176

Saturation mapping with subclones of YACs: DNA marker production targeting the rice blast disease resistance gene, Pi-b

  • L. Monna
  • A. Miyao
  • H. S. Zhong
  • M. Yano
  • M. Iwamoto
  • Y. Umehara
  • N. Kurata
  • H. Hayasaka
  • T. Sasaki

DOI: 10.1007/s001220050396

Cite this article as:
Monna, L., Miyao, A., Zhong, H. et al. Theor Appl Genet (1997) 94: 170. doi:10.1007/s001220050396

Abstract

Saturation mapping of a very small genomic region is indispensable for map-based cloning. We applied a method based on sub-cloning and the Southern-hybridization technique for generating RFLP markers directly from yeast artificial chromosomes (YACs). Two YACs overlapping each other and covering the locus of the rice blast resistance gene, Pi-b, were used to construct a plasmid sub-library. Rice-specific and single-copy clones suitable as probes for RFLP analysis were selected from this sub-library by hybridization to the blots of digested DNAs of rice, YACs, and yeast. As a result, 22 markers were produced within a small chromosomal region including Pi-b. This case study shows that overlapping YACs known to cover the gene of interest are very useful in fine-scale physical mapping leading to map-based cloning of the target gene.

Key words Saturation mappingYeast artificial chromosome (YAC)RicePi-bBlast disease resistance

Copyright information

© Springer-Verlag Berlin Heidelberg 1997

Authors and Affiliations

  • L. Monna
    • 1
  • A. Miyao
    • 1
  • H. S. Zhong
    • 1
  • M. Yano
    • 1
  • M. Iwamoto
    • 1
  • Y. Umehara
    • 1
  • N. Kurata
    • 1
  • H. Hayasaka
    • 2
  • T. Sasaki
    • 1
  1. 1.Rice Genome Research Program, National Institute of Agrobiological Resources and Institute of Techno-innovation of Agriculture, Forestry and Fisheries, 446-1 Ippaizuka, Kamiyokoba, Tsukuba, Ibaraki, JapanJP
  2. 2.Miyagi Prefectural Furukawa Agricultural Experiment Station, Suwa, Furukawa, Miyagi, JapanJP