| Project/Area Number |
11660076
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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 |
応用微生物学・応用生物化学
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| Research Institution | Tokyo University of Agriculture and Technology |
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Principal Investigator |
KAWAI Shinya Faculty of Agriculture, Tokyo University of Agriculture and Technology Associated professor, 農学部, 助教授 (90202027)
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| Project Period (FY) |
1999 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2000: ¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1999: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Keywords | lignin / vascular bundle / transcriptional factor / Lim protein / aspen / リグニン / 樹木 / プロモーター |
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| Research Abstract |
We already isolated many lignin biosynthetic genes from hybrid aspen. And genes which expressed in vascular bundle were specified from them. Using promoter regions of them, we aimed to isolate transcriptional factors which control gene expression in vascular bundle of woody plants. Promoter regions of pal g2b and prxA3a, which specifically related to lignin biosynthesis, were respectively shortened to about 200 bps, in which cis elements existed that controlled gene expression in vascular bundle. There were cis element motifs to which Lim-like transcriptional factors bind. Ntlim1 cDNA was already isolated from Nicotiana tabacum, which encoded Lim-like protein, and it is known that down- regulation of NtLim1 caused inhibition of lignin accumulation in tobacco and eucalyptus plants. We isolated cDNA 98% identical to Ntlim1from N.tabacum SumSunNN by using RT- PCR techniques. It was thought to be a variation among strains. And it was expressed in E.coli as 6 x His tag fused protein. It was purified and used in gel mobility shift assay. But it aggregated and didn't attach to FITC labeled probes. We also isolated cDNA encoding a protein about 90% identical to NtLim1 protein. And its promoter region was amplified and isolated by using inverse PCR techniques.
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