| Project/Area Number |
01400001
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| Research Category |
Grant-in-Aid for General Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Research Field |
広領域
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| Research Institution | Tsukuba University |
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Principal Investigator |
FUJII Tadashi University of Tsukuba Institute of Biological Sciences Professor, 生物科学系, 教授 (20011611)
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| Co-Investigator(Kenkyū-buntansha) |
HARA Yukichi 東京医科歯科大, 医学部, 助教授 (00092429)
SATOH Shinobu University of Tsukuba Institute of Biological Sciences Assistant Professor, 生物科学系, 講師 (70196236)
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| Project Period (FY) |
1989 – 1990
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| Project Status |
Completed (Fiscal Year 1990)
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| Budget Amount *help |
¥16,600,000 (Direct Cost: ¥16,600,000)
Fiscal Year 1990: ¥7,000,000 (Direct Cost: ¥7,000,000)
Fiscal Year 1989: ¥9,600,000 (Direct Cost: ¥9,600,000)
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| Keywords | Raphidophyceae / Na^+-ATPase / halo-tolerant / ラフィド藻 / イオン輸送 / Heterosigma akashiwo / Na^<++>-activated ATPase / PCR(ポリメラ-ゼ・チェィン・リアクション) / cDNA |
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| Research Abstract |
The ability which can export intracellular sodium ions can be transformed usual higher plants to halotolerant. The Na^+ K^+-ATPase of animal cells or Na^+-ATPase of raphidophycean H. akashiwo was able to transport Na ions. The Na^+-ATPase of H. akasiwo was consequently proper enzyme for the transformation. A DNA between conserved parts of eucaryotic ATPase was amplified from H. akasiwo cDNA library with PCR method. The Na^+-ATPase of H. akashiwo was cloned with the amplified DNA as a probe. Two kinds of ATPase molecules was cloned consequently. The amino acid sequences and the secondary structures analysis showed the homology to other species' ATPases(70% : protozoan L. donovani, 40-50% : higer plants and fungi, 20-30% : higher animals). This suggests that the ATPase amino acid sequences tend to conserve among systematical close to species rather than the functional characters (ion selectivity, etc). Rescreeing was needed to the reason why the two cDNA clones were short of complete coding regions. The most expressing cell period of the genes was determined with northern hybridization and 5' end probe was successed to get the enough length cDNA (3.4kb).
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