1993 Fiscal Year Final Research Report Summary
Development and application of new regulatory genes
| Project/Area Number |
02403023
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| Research Category |
Grant-in-Aid for General Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Research Field |
応用生物化学・栄養化学
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| Research Institution | Kyoto University |
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Principal Investigator |
KOMANO Tohru Kyoto University Faculty of Agriculture Professor, 農学部, 教授 (30026413)
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| Co-Investigator(Kenkyū-buntansha) |
KIOKA Noriyuki Kyoto University Faculty of Agriculture Assistant Professor, 農学部, 助手 (90234179)
UEDA Kazumitsu Kyoto University Faculty of Agriculture Assistant Professor, 農学部, 助手 (10151789)
SAKAI Hiroshi Kyoto University Faculty of Agriculture Associate Professor, 農学部, 助教授 (60089117)
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| Project Period (FY) |
1990 – 1993
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| Keywords | Replication initiation region / Promoter of BT / SSI signal / Heat shock response / Multi-drug resistance / Glyco-technology |
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| Research Abstract |
Two ssi signals essential for the initiation of a broad hostrange plasmid RSF1010 were identified. Replication of RSF1010 required plasmid-encoded RepC initiator protein. The RepC was able to bind specifically to a cluster of the three and a half iterons contained in oriVRSF1010. A single 20-bp iteron itself was sufficient for the RepC binding, while it was neither able to support the plasmid replication nor to exert incompatibility. Treatment of the RepC dimers with chemical denaturants was shown to activate RepC binding to the iterons. The DnaK heat shock protein was proved essential for the RSF1010 replication in vivo. DnaK playd a major role in the RepC activation process.
The cryIVB gene of Bacillus thuringiensis subsp. israelensis coded insecticidal protein. Thepromoter of this gene was confirmed to bind only alpha^<35>, which controled this gene expression at the midsporulation stage.
In a well defined cell line HepG2, sodium arsenite treatment led to a 2-3 fold increase of MDR gene expression and Pglycoprotein synthesis. Transcriptional activation after exposure to arsenite depends on a 60-bp region containing two heat-shock responsive elements. The increase of P-glycoprotein synthesis and MDRI mRNA caused by arsenite was inhibited by quercetin. A transcellular transport system revealed some anti-cancer drugs were transported through binding of these drugs to Pglycoprotein.
N-acetylglucosaminyltransferaseI(GnT-1)cDNA was isolated and expression in several rat tissues was analyzed. GnT-1 mRNA was expressed in a tisseue specific manner and the pattern of the expression was species specific.
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