Saturation mapping with subclones of YACs: DNA marker production targeting the rice blast disease resistance gene, Pi-b |
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| Authors: | L Monna A Miyao H S Zhong M Yano M Iwamoto Y Umehara N Kurata H Hayasaka T Sasaki |
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| Institution: | (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, Japan, JP;(2) Miyagi Prefectural Furukawa Agricultural Experiment Station, Suwa, Furukawa, Miyagi, Japan, JP |
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| 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.
Received: 2 May 1996 / Accepted: 2 August 1996 |
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| Keywords: | Saturation mapping Yeast artificial chromosome (YAC) Rice Pi-b Blast disease resistance |
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