Project/Area Number |
07660030
|
Research Category |
Grant-in-Aid for Scientific Research (C)
|
Allocation Type | Single-year Grants |
Section | 一般 |
Research Field |
園芸・造園学
|
Research Institution | Faculty of Agriculture, Shinshu University |
Principal Investigator |
OHI Michio Shinshu Unv., Fac.of Agric., assistant Professor, 農学部, 助教授 (80167296)
|
Project Period (FY) |
1995 – 1997
|
Project Status |
Completed (Fiscal Year 1997)
|
Budget Amount *help |
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1997: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1996: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1995: ¥1,100,000 (Direct Cost: ¥1,100,000)
|
Keywords | Allium grayi / FISH / Sterile strain / PCR / Chromosomes / cDNAライブラリー / In situハイブリダイゼーション / FISH法 |
Research Abstract |
Deference of chromosomes constitution between fertile strain and sterile strain in Allium grayi was researched by in situ hybridization. Probes for FISH were produced from genomic DNA in A.grayi.. The genomic DNAs extracted from young leaves were amplified by PCR using forty primers (Operon 10-mer Kits, A Kit 1-20, B Kit 1-20). From the result, Polymorphism's between two strains appeared in twenty primers. The polymorphic bands by electrophoresis were cut out and refined. These DNA fragments were amplified by PCR using the same probes. Twelve probes were retrieved by electrophoresis from 400 bp to 1800 bp. Obtained probes were screening by southern hybridization using ECL system of Amersham. However, No probes which were identified as fluorescent signals on the nitrocellulose filter. We are, presently, attempting selection of probes radiated and trying in situ PCR method, amplify the probes directly on the sample of somatic chromosomes.
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