1999 Fiscal Year Final Research Report Summary
Development of a new differential display and its application
| Project/Area Number |
09044273
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| Research Category |
Grant-in-Aid for Scientific Research (A).
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Human genetics
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| Research Institution | Tokyo Medical and Dental University, Medical Research Intitute |
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Principal Investigator |
YASUKOCHI Yukio Tokyo Medical and Dental University, Medical Research Institute, Professor, 難治疾患研究所, 教授 (60037398)
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| Co-Investigator(Kenkyū-buntansha) |
WATANABE Teruo University of Tsukuba School of Medicine, Professor, 医学専門群, 教授 (40037396)
TAUCHIYA Terumasa Medical and Dental University, Medical Research Institute, Assistant Professor, 難治疾患研究所, 助手 (20242109)
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| Project Period (FY) |
1997 – 1999
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| Keywords | Gene expression / Human fetal brain / Differential display / Protein tyrosine kinase |
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| Research Abstract |
1. Temporary expression of A31 mRNA in nascent dendrites of human fetal brain.
A differential display method was used to compare patterns of mRNA expression between human fetal and adult brains and we obtained 26 candidates for highly expressed fetal cDNA fragments. A clone, A31, had no significant homology in sequence compared with the DDBJ/EMBL/GenBank databases. In the RT-PCR and Northern analyses, the mRNA was about 9kb and only expressed in middle state of human fetal brain, and absent in various tissues except brain. Further characterization of this clone was performed by in situ hybridization, which showed that the mRNA was mostly expressed in cell bodies and dendrites of neurons in cortical plates. Our analyses suggest that this mRNA is involved in dendritic development at the middle stage in human fetal brain.
2. A method for differential display of protein tyrosine kinase cDNAs from the human fetal and adult cerebral brains.
We describe a differential display for profiling the expression of protein tyrosine kinases and apply it to cDNAs from human fetal and adult cerebral brains. The method involves two selective steps for processing the mRNA.At each step degenerate oligonucleotide primers derived from highly conserved regions of the catalytic domain of the kinases are used. In the display with BstY I and BsiHKA I digests of the cDNA, 65% and 59% of a total of 72 and 63 bands, respectively, represented fragments from a total of 27 different kinases. The expression levels of the kinases in the diplay were comparable with those measured by RT-PCR.The present method offers a relatively specific way to display differentially expressed gene families in any tissues and cell type.
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Research Products
(2 results)
