2001 Fiscal Year Final Research Report Summary
The expression analysis of symbiotic nitrogen-carbon
| Project/Area Number |
12660061
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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 |
Plant nutrition/Soil science
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| Research Institution | Kagawa University |
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Principal Investigator |
TAJIMA Shigeyuki Kagawa University, Faculty of Agriculture, Professor, 農学部, 教授 (50116894)
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| Co-Investigator(Kenkyū-buntansha) |
AGARIE-NOMURA Mika Kagawa University, Faculty of Agriculture, Assistant Professor, 農学部, 助手 (50294749)
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| Project Period (FY) |
2000 – 2001
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| Keywords | metabolism / transgenic plant / symbiotic nitrogen fixation / cloning / uricase / high level expression / promoter analysis / gel shift assay |
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| Research Abstract |
We have cloned and sequenced uricase genes of various legume plants and made a phylogenetic tree. As a result, phylogenetic analysis of uricase sequences from various legumes indicated that uricases of amide- and ureide- transporting legumes forms two distinct groups, suggesting these types of legumes were evolutionary divided.
The two uricase genes of soybean were expressed at high level using pET vector. After purification of two uricases, we investigated these uricase characterizations. There was no different Km value against oxygen between them. However, one uricase (UR9) showed lower Km value against uric acid than another uricase (UR2).
Gain of function and loss of function experiments suggested that 6 base (GTAATG) motif in the 5' non-coding region of soybean uricase gene was a cis element for nodule uninfected cell specific expression.
We produced the transgenic lotus plants for high level expression of PEPC or PPDK to analyze C or N metabolism in nodule.
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