| Project/Area Number |
09306006
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| Research Category |
Grant-in-Aid for Scientific Research (A).
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
応用微生物学・応用生物化学
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| Research Institution | KYOTO UNIVERSITY |
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Principal Investigator |
OHYAMA Kanji Kyoto Univ., Grad. Sch. Biostudies, Professor, 生命科学研究科, 教授 (40135546)
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| Co-Investigator(Kenkyū-buntansha) |
YAMATO Katsuyuki Kyoto Univ., Grad. Sch. Biostudies, Assistant, 生命科学研究科, 助手 (50293915)
FUKUZAWA Hideya Kyoto Univ., Grad. Sch. Biostudies, Associate professor, 生命科学研究科, 助教授 (30183924)
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| Project Period (FY) |
1997 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥33,300,000 (Direct Cost: ¥33,300,000)
Fiscal Year 2000: ¥4,000,000 (Direct Cost: ¥4,000,000)
Fiscal Year 1999: ¥4,700,000 (Direct Cost: ¥4,700,000)
Fiscal Year 1998: ¥8,200,000 (Direct Cost: ¥8,200,000)
Fiscal Year 1997: ¥16,400,000 (Direct Cost: ¥16,400,000)
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| Keywords | Liverwort / sex chromosomes / PAC libraries / EST / sex-chromosome genes / RDA / transformation / FISH / Y染色体 / PACクローン / 反復配列 / PACライブラリー / 生殖器官 / PAC |
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| Research Abstract |
We constructed PAC genomic libraries for both male and female liverwort, Marchantia polymorpha. A male-specific clone, pMM4G7, was isolated from the male genomic library by differential screening and shown to originate from the Y chromosome by FISH.The entire nucleotide sequence of pMM4G7 was determined to find Y-chromosome-specific repeated sequences and a gene coding for a protein with a RING-finger motif. Another sequenced male-specific PAC clone, pMM2D3, containd six genes which showed significant similarities to genes registered in the public databases. Four of the six genes were present only in the male genome, and RT-PCR showed that these were actually expressed. Five novel markers for the Y chromosome and two for the X chromosome were isolated by representational difference analysis (RDA). Total sizes of contigs constructed thus far are approximately 3Mb and 150kb for the Y and X chromosomes, respectively.
Separate cDNA libraries were constructed for male and female sexual organs. Approximately 1000 ESTs were obtained from each of the libraries, revealing gene expression profiles in sexual organs as well as differences in the expression profiles between male and female sexual organs.
In order to elucidate functions of novel genes encoded by the sex chromosomes by gene manipulation, we developed a transformation system for M.polymorpha applying particle bombardment. We have constructed a library of over 2000 tagged lines of M.polymorpha and isolated a mutant which was affected in the development of sexual organs.
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