| Project/Area Number |
61480156
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Immunology
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| Research Institution | Cancer Institute, Kanazawa University |
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Principal Investigator |
TAKAHASHI Morinobu Cancer Institute, Kanazawa University, がん研究所, 教授 (80019877)
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| Project Period (FY) |
1986 – 1987
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| Project Status |
Completed (Fiscal Year 1987)
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| Budget Amount *help |
¥7,000,000 (Direct Cost: ¥7,000,000)
Fiscal Year 1987: ¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1986: ¥4,000,000 (Direct Cost: ¥4,000,000)
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| Keywords | MHC class III / Complement C4 / Sex-limited protein / Transcriptional regulation / 転写調節 / CTIアツセイ / マウス補体C4 / sex-limited Protein / テストステロン / 遺伝子発現 |
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| Research Abstract |
Mouse C4 and Sex-limited protein(Slp) are encoded by two recently duplicated genes of the major histocompatibility complex III of the mouse. The availability of many expression variants for these genes among mouse strains makes them the attractive system in which the regulation of eukaryotic genes under developmental, hormonal and tissue specific control can be studied. We isolated and characterized cDNA and genomic DNA clones for these genes from several mouse strains which show interesting variation for the expression of C4 and slp. Our experimental results obtained by various molecular genetic techniques can be summarized as follows.
1. The mouse strains carrying H-2^k haplotype are characterized as low C4 producers. We measured C4 mRNA by Northern blotting and nuclear C4 RNA by nuclear run on assay using the hepatocytes from high C4 producing mouse(B10) and low C4 producing mouse(B10.BR). Furthermore we tested the 5'flanking regions of the C4 genes from these strains for their trans
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criptional regulatory activity with transfected cells. The results clearly showed that the low C4 bisynthesis is mainly controlled at the posttranscriptional and pretranslational level.
2. To elucidate the molecular basis underlying the difference in the mode of expression of C4 gene(constitutional) and Slp gene(testosterone induced), we compared the nucleotide sequences of these two genes isolated from FM strain. Furthermore we tested the 5'flanking region fragments for transcriptional regulatory cativity using CAT assay in transfected cells. We identified the functional domains in the regions of these genes that appear to explain the difference in the gene expression of the C4 and Slp genes.
3. We isolated and characterized all of C4-related genes from the cosmid library of C3H.W7 strain that shows an extraordinary mode of expression of Slp(constitutional expression). We found the three of apparent Slp genes indeed consisting of the C4-derived 5' half and Slp-derived 3' half. By comparing their nucleotide seguences with those of C4 and Slp genes, we concluded that these recombinant genes are derived by multiple cross over in the central portions of the gene. Less
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