1999 Fiscal Year Final Research Report Summary
Mapping and cloning of the Da mutant gene defective dorsal-ventral patterning
| Project/Area Number |
10680686
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Developmental biology
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| Research Institution | NAGOYA UNIVERSITY |
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Principal Investigator |
OZATO Kenjiro Nagoya University, Bioscience Center, Profesor, 生物分子応答研究センター, 教授 (90026790)
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| Co-Investigator(Kenkyū-buntansha) |
KIMURA Minoru Tokai University, Medical School, Professor, 医学部, 教授 (10146706)
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| Project Period (FY) |
1998 – 1999
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| Keywords | medaka / morphogenesis / dorsal-ventral patterning / positional cloning |
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| Research Abstract |
Formation of dorsal-ventral patterning is one of the major issues of vertebrate development. Medaka Da mutant expresses a phenotype in which structures characteristic to the dorsal region are ventralized. In the present study, we attempted to identify the Da gene by positional cloning.
1) Construction of the linkage map and mapping of the Da gene.
The genetic markers we used allowed us to identify 26 linkage groups in the medaka genome, in which the Da gene was mapped on the linkage group VIII. The physical distance from Da to two markers between which put Da was 144kb and 360kb, respectively. These distances were close enough to conduct chromosomal walking.
2) Construction of Cosmid library
We constructed a Cosmid library using the medaka HNI-I strain, which consisted of 120,000 clones (corresponding to 6 haploid genomes) with an average insert-length of 40.2kb. We successfully obtained Cosmid clones containing two markers close to Da by screening the library.
3) Identification of Da in one BAC clone.
Using Cosmid and BAC clones, we determined a minimum region containing Da, demonstrating that Da was contained in a range of 70kb to 250kb. Thus, it was expected that the Da gene was covered by one BAC clone.
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