| Project/Area Number |
09660357
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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 |
Applied molecular and cellular biology
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| Research Institution | KYOTO UNIVERSITY |
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Principal Investigator |
FUKUZAWA Hideya Kyoto University, Graduated School of Agriculture, Associate Professor, 農学研究科, 助教授 (30183924)
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| Project Period (FY) |
1997 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 1998: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1997: ¥1,700,000 (Direct Cost: ¥1,700,000)
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| Keywords | Chlamydomonas / carbonic nahydrase / carbon concentrating mechanisms / signal transduction / photosynthesis / carbon dioxyde / stress response / レポーター遺伝子 / アリルスルファターゼ / 炭素脱水酵素 / 緑藻クラミドモナス |
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| Research Abstract |
Several chimeric constructs were generated by fusing series of 5'-upstream region of CAHl to the arylsulfatase reporter gene (ars) driven by βィイD22ィエD2-tublin minimal promoter. Some Chlamydomonas cells transformed with the chimeric constructs expressed the reporter gene in response to changes in the external COィイD22ィエD2 concentration. The 5'-upstream region is divided into two parts ; a part functions as a silencer in high-COィイD22ィエD2 condition and the other part functions as an enhancer in low-COィイD22ィエD2 condition.
Seven high-COィイD22ィエD2-requiring mutants, which cannot grow as wild type Chlamydomonas does in air level of COィイD22ィエD2(0.04%), were isolated by gene tagging. It was indicated that these mutants carried lesions in genes related to the carbon concentrating mechanism (CCM) or the signal transduction pathway between sensing changes of environmental COィイD22ィエD2 concentration and induction of CCM-related genes. By using an indexed PAC genomic library, two mutants were complemented. The mutation of a mutant C1, which accumulate less inorganic carbon (Ci) into the cell, was complemented by a PAC clone containing 20-kb of genomic DNA. This suggests that Ci-transport related genes could be encoded within this DNA fragment. Another mutant C16, which does not induce CCM-related genes in low-COィイD22ィエD2, was transformed by a DNA fragment which correspond to the flanking sequences of the NIT1 insertional locus and the transformants recovered the induction of CCM-related genes. The cloned DNA fragment encoded a gene which does not show significant homology with any previously known genes.
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