Project/Area Number |
01560093
|
Research Category |
Grant-in-Aid for General Scientific Research (C)
|
Allocation Type | Single-year Grants |
Research Field |
応用生物化学・栄養化学
|
Research Institution | Nagoya University |
Principal Investigator |
ASAHI Tadashi Nagoya University School of Agriculture Professor, 農学部, 教授 (10023392)
|
Co-Investigator(Kenkyū-buntansha) |
NAKAMURA Kenzo Nagoya Univ., School of Agric., Associate Professor, 農学部, 助教授 (80164292)
|
Project Period (FY) |
1989 – 1990
|
Project Status |
Completed (Fiscal Year 1990)
|
Budget Amount *help |
¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1990: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1989: ¥1,300,000 (Direct Cost: ¥1,300,000)
|
Keywords | Castor bean / Catalase gene / Plant catalase / Promotor / Regulation of gene expression / Transgenic plants / 遺伝子発現調節 / 種子発芽 / 融合遺伝子 |
Research Abstract |
There are several catalase genes in the castor bean genome : two of them (CAT1 and CAT3 genes) have been cloned, and it has been observed that CAT1 gene expresses only in the endosperm and cotyledons during seed germination, whereas CAT3 gene does in all tissues of the seedlings. In the present study, the structures of the two genes were completely determined and CAT3 gene was found not to express in the green leaves. In order to analyze the molecular mechanism of regulation of the gene expression, fused genes of 5'-upstream regions with various lengths of the genes with Escherichia coli beta -glucuronedase gene (CAT1-GUS and CAT3-GUS fused genes) were constructed and their transient or stable expression in tobacco cultured cells was analyzed. Long 5'-upstream regions of both genes showed strong promotor activities in either expression ; especially the regions of CAT3 gene showed stronger activigties than the promotor of cauliflower mosaic virus 35S RNA gene, which has been used for strong expression of foreign genes in transgenic plants. The stable expression analysis suggests that there are many cis-elements enhancing the promotor activity in the 5'-upstream regio of CAT3 gene, whereas only a few elements exists in the region of CAT1 gene. In transgenic tobacco plants carrying CAT3-GUS fused gene, the foreign gene expressed in all types of cells except for the fused genes of very short 5'-upstream regions which showed tissue-specific expression. This suggests the presence of several cis-elements concerning tissue-specific expression in the long 5'-upstream region.
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