| Project/Area Number |
08671324
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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 |
Embryonic/Neonatal medicine
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| Research Institution | Fujita Health University |
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Principal Investigator |
YAMADA Kazuyo Institute for Comprehensive Medical Science, Fujita Health University lecturer, 総合医科学研究所, 講師 (90080217)
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| Co-Investigator(Kenkyū-buntansha) |
HONDA Shin-ichiroh Institute for Comprehensive Medical Science, Fujita Health University Research a, 総合医科学研究所, 助手 (40257639)
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| Project Period (FY) |
1996 – 1997
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| Project Status |
Completed (Fiscal Year 1997)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1997: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1996: ¥1,300,000 (Direct Cost: ¥1,300,000)
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| Keywords | transcription factor / aromatase / trophoblast / placenta / 栄養胚芽細胞 / hCG / プロモーター / シスエレメント |
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| Research Abstract |
We found a new cis-element neccessary for the placenta-specific enhancer in the aromatase gene. Co-presence of the element and the trophoblast specific element (TSE) in the aromatase enhancer region is sufficient to direct trophoblast specific expression driven by a heterologous tk promoter. The binding activity to this site was found in the nuclear extracts prepared from JEG-3 cells and the human placenta but not in the cell extract from HeLa or Hep G2 cells. This binding activity is distinct in electrophoreic mobility from the protein that binds to the TSE-like elements in aromatase enhancer. The core DNA sequence for the binding of the new trans-factor is also distinct from that of the TSE binding protein. The similar sequence is however found in the 24-base pair Trophoblast Specific Element (TSE) derived from the a-hCG promoter in reverse direction thus this DNA fragment also binds to the new factor with low affinity. This new trans-factor is tentatively called TSEBP2.
A2.65 Kilobase cDNA clone that encodes a 436 amino acid was isolated from a human placental cDNA library using a yeast one-hybrid system with the TSEBP2 recognition sequence (GATCCATAAGACCCTCATTCCAGAG). The transfection of this clone with baculovirus system in sf21 cells resulted the production of a nuclear protein that showed the same binding specificity with the TSEBP2 in the nuclear extract prepared from Jeg 3. Northern-blot analysis showed that the mRNA coresponding to this clone was found exclusively in the placenta. This protein is likely to be a trophoblast specific transcription factor that confer placenta-specific expression of aromatase gene. The primary structure of the tentative transactivation domain however, has no similarity with other known transcription factors. The proof of transcriptional activity is on the way.
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