Development of systems evaluating evolution at gene expression level
| Project/Area Number |
06554038
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| Research Category |
Grant-in-Aid for Developmental Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
系統・分類
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| Research Institution | University of Tokyo |
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Principal Investigator |
UEDA Shintaroh University of Tokyo, Graduate School of Science Associate Professor, 大学院・理学系研究科, 助教授 (20143357)
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| Co-Investigator(Kenkyū-buntansha) |
SAITOU Naruya National Institute of Genetics Associate Professor, 助教授 (30192587)
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| Project Period (FY) |
1994 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥6,300,000 (Direct Cost: ¥6,300,000)
Fiscal Year 1995: ¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1994: ¥4,500,000 (Direct Cost: ¥4,500,000)
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| Keywords | gene expression / transcription factor / target DNAs / molecular evolution / 種 / 形態進化 / 遺伝子発現系 |
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| Research Abstract |
There have been reported a number of studies on molecular evolution since 1960s, but most of them are studies on phylogenetic relationships among organisms. It is now clear that it is hard to deduce answer about evolutionary mechanisms at molecular level from these molecular phylogenetic studies. Then, we tried here to develop systems for evaluating evolution at gene expression level by using a POU transcription factor as a model case. First, we constructed an expression vector involving DNA-binding domain of POU,transformed it into Escherichia coli, deduced specifically POU protein, and purified it by affinity chromotography. Using POU protein purified, we tried to separate target DNAs of POU,based on affinity between POU protein that binds its target DNAs and antibodies against POU protein (antibody method) or on DNA-protein binding between POU and its target DNAs (DNA-binding method). We separated several candidate DNAs (genomic DNAs) using DNA-binding method. We then screened brain cDNA library using these DNAs as probes, and obtained two independent cDNA clones of target. Nucleotide sequences and their deduced amino acid sequences showed no sequence homology to those so far reported. We furthermore developed evaluating system for gene expression using POU transcription factor and its target sequences obtained in this study, which based on yeast GAL4 system. This system would be available for all of eukaryote species excluding yeast in examining evolutionary mechanisms at molecular level, by slight modification of this system onto cassette system where multiple restriction enzymes are available.
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Report
(3 results)
Research Products
(10 results)
