| Project/Area Number |
14580713
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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 |
Developmental biology
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| Research Institution | Nara Women's University, Graduate School of Natural Science and Ecological Awareness, Department of Biological Science and Environment |
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Principal Investigator |
HARUMOTO Terue Nara Women's University, Graduate School of Natural Science and Ecological Awareness, Department of Biological Science and Environment, 大学院・人間文化研究科, 助教授 (80198936)
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| Co-Investigator(Kenkyū-buntansha) |
IIO Hideo Osaka City University, Professor, 大学院・理学研究科, 教授 (80145771)
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| Project Period (FY) |
2002 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 2004: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 2003: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2002: ¥1,700,000 (Direct Cost: ¥1,700,000)
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| Keywords | Blepharisma / conjugation / ciliate / PCR / regulation of expression / gene / promoter / Cdk / ガモン / 転写調節因子 / サブトラクション / 未熟期 |
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| Research Abstract |
This study deals with the molecular mechanism of induction of conjugation in dliates, especially in heterotrich ciliate Blepharisma japonicum.
1.Regulation of expression of gamone 1 gene in type I cells.
We investigated the condition which ensures the gamone 1 expression. We revealed that gamone 1 gene is restrictively transcribed in mature type I cells under appropriate starved condition. The transcription was promoted by complementary gamone 2 and the level of starvation.
2.Sequence determination of genomic gamone 1 gene in type I and type II cells.
To analyze the regulation of gene expression of gamone 1 gene, we cloned and sequenced the genomic sequence of gamone 1 gene in type I and type II cells. The study revealed that intron was not included in the genomic gamone 1 gene in both type I and type II cells. The sequence of the gamone 1 gene in type n cells coincides with that of in type I cells.
3.Analysis of the regulation of gene expression of gamone 1 gene encoding conjugation-inducing substance.
Using inverse PCR technique, we cloned and sequenced 868 bp of the upstream of gamone 1 gene in type I cells. We found two TATA boxes, a CCAAT box, a PSE (proximal sequence element) and several sites which are recognized by known transcriptional factors.
4.Identification of genes which are specifically expressed in conjugation-induced cells.
Several genes are considered to be activated in conjugation-induced type II cells. We subtracted expressed genes between gamone-treated and untreated cells, and identified several genes which were specifically expressed in gamone-treated cells. We found genes encoding Cdk, Cks and 4-HPPD. These genes were transcribed in 2-4 hours after induction of conjugation.
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