| Project/Area Number |
16590822
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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 |
Neurology
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| Research Institution | Faculty of Nursing, Shiga University of Medical Science |
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Principal Investigator |
YASUDA Hitoshi Shiga University of Medical Science, Faculty of Nursing, Professor, 医学部, 教授 (80135467)
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| Co-Investigator(Kenkyū-buntansha) |
KOJIMA Hideto Shiga University of Medical Science, Dept Molecular Genetics in Medicine, Associate Professor, 医学部, 助教授 (00225434)
MAEDA Kengo Shiga University of Medical Science, Devision of Neurology, Department of Medicine, Assistant Professor, 医学部, 助手 (80324581)
KAWAI Hiromichi Shiga University of Medical Science, Devision of Neurology, Department of Medicine, Assistant Professor, 医学部, 助手 (90378456)
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| Project Period (FY) |
2004 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2005: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2004: ¥2,500,000 (Direct Cost: ¥2,500,000)
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| Keywords | Dorsal Root ganglion neurons / Phage display / Nervre growth factor / Erythropoietin / Adeno-associated virus / CMV promotor / 脊髄前角細胞 / 後根神経節ニューロン / phage display peptide library / 大腸菌 / プラーク数 / DNA sequence |
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| Research Abstract |
Dorsal root ganglion (DRG) neurons isolated from adult C57/BL mouse was placed with phage samples from phage display peptide library (Ph.D. -CX7C kit) including huge numbers of polypeptides with specific arrangement (7 amino acids) for 10 min. The collected phage was infected by E.coli, thereby formed plaques overnight. DNA sequence was obtained from the phages forming plaques, thereby providing 3 kinds of peptide arrangements composed of 7 amino acids which are supposed to show a strong affinity to DRG neurons. In fact, these peptides were confirmed to show an immunoreactivity with DRG neurons using anti-phage antibody. In spite of successful progress, the experiment had to be discontinued because it was announced on the 29 th of January, 2004, that the research which uses phage has come to require the permission from the Ministry of Education, Culture, Sports, Science and Technology. Because it was expected that it took considerable time until getting the permission, in place of phage, adeno-associated virus (AAV) which has a affinity to DRG neurons was used as vector for a while. Shuttle vector was made by connecting CMV promoter and polyA to nerve growth factor and erythropoietin genes which were made by PCR method. Since these genes were somewhat small for connecting to AAV vector, stuffer was used. On the 6^<th> of February, 2006, the announcement notifying that we are able to do experiment without specific permission was from the government. Then, we restarted the experiment along the original line using the phage as vector. Because of the above interruption of experiment during one year, we have not obtained the expected progress. However, we have obtained 3 kinds of the peptides specific for DRG neurons. These peptides would provide powerful therapeutic methods for neurodegenerative disorders including amyotrophic lateral sclerosis.
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