2006 Fiscal Year Final Research Report Summary
Development of novel mutant screening system with yeast homologus recombination and it's reverse genetic applications.
| Project/Area Number |
17580008
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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 |
Breeding science
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| Research Institution | Saga University |
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Principal Investigator |
ANAI Toyoaki Saga University, Faculty of Agriculture, Associate Professor, 農学部, 助教授 (70261774)
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| Co-Investigator(Kenkyū-buntansha) |
NAGANO Yukio Saga University, Analytical Research Center for Experimental Sciences, Associate Professor, 総合分析実験センター, 助教授 (00263038)
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| Project Period (FY) |
2005 – 2006
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| Keywords | soybean / mutant / reverse genetics |
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| Research Abstract |
In recent years, genomic information containing genomic and EST sequences of various organisms is rapidly accumulating in public database. The huge nucleotide sequence data may be valuable for elucidating the molecular basis of beneficial genes in crop improvement. However, it is quite difficult to apply the map-based analysis methods because of almost all crop plants carrying the highly complicated genomes. Hence, novel robust reverse genetics-based tools are essential to link between the huge nucleotide sequence data and specific gene function.
In this study, we have developed the soybean mutant library consisting of M2 genomic DNA stocks and M3 seed stocks of about 28 thousand individual mutant lines with following procedure.
1. Soybean mutant population (Ml) was developed by X-ray (200Gy) irradiation to dry seeds.
2. Self-pollinated M2 seeds were grown in the field, and green leaves and matured M3 seeds were harvested from individual plants.
3. After extraction of genomic DNA from M2 individuals, and they randomly amplified with the multiple displacement amplification (MDA) method. Phi29 DNA polymerase, the key enzyme of MDA reaction, was expressed in E. coli system and purified with a chitin-based affinity chromatography.
We successfully obtained soybean mutant library, however, further improvement will be needed to establish the yeast-based mutant detection system.
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Research Products
(4 results)
