2005 Fiscal Year Final Research Report Summary
Analysis of indican synthesis pathway in Indigo plant (Polygonaceae).
| Project/Area Number |
14540605
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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 |
植物生理
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| Research Institution | Okayama university of science |
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Principal Investigator |
MINAMI Yoshiko Okayama university of science, Biochemistry, Assistant professor, 理学部, 助教授 (60192762)
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| Project Period (FY) |
2002 – 2005
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| Keywords | Secondary metabolism / Indican / Indigo / Tryptophan synthase / UDP-glucosyl transferase / cDNA cloning / Cytochrome P450 |
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| Research Abstract |
Indican, one of the secondary metabolites, is stored in the leaves of Indigo plant (Polygonaceae). Probably, indican is synthesized from indole, an intermediate of tryptophan synthesis pathway. Indole is oxidized to indoxyl by cytochrome P450. Furthermore, indoxyl is converted to indican by UDP-glucosyltransferase. I predicted the presence of switching mechanism between tryptophan and indican synthesis pathways.
1.cDNA cloning of UDP-glucosyltransferase
A desired clone could not be gotten, although cDNA cloning was tried by some methods. By two-dimensional electrophoresis, the total proteins of callus were compared with those of leaves, the difference was analyzed. Still now, we continue the analysis.
2.cDNA cloning and analysis of cytochrome P450
ptP450 cDNA was isolated from cDNA library of indigo plant. The amino acid sequence of ptP450 is homologous to those of P450s, which react with some alkaloids including indole. Therefore, I tried the analysis of properties of recombinant ptP450 that was expressed in E.coll.
3.cDNA cloning and analysis of tryptophan synthase
Three cDNAs of tryptophan synthase β were amplified by RT-PCR. Now we examine the difference of expression by Northern blot.
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