Immunohistochemical Detection of Human Cellular Yes Gene (C-Yes-1) Messenger Rna by A Non-Radioactive Synthesized Oligodeoxynucleotide Probe
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概要
- 論文の詳細を見る
A synthetic 39-mer anti-sense strand oligodeoxynucleotide complementary to the region of human cellular yes gene (c-yes-1) mRNA that codes for a polypeptide of 543 amino acids (Mr: 60, 801) and the corresponding sense strand 39-mer oligodeoxynucleotide were chemically synthesized by β-cyanoethyl phospho-ramidite method. The probes thymine-thymine (T-T) dimerized by UV irradiation were used for in situ hybridization studies. Various cells were fixed with ethanol-acetic acid (3:1) at room temperature for 20min and hybridization was carried out in 50% formamide for 36hr at 37°C. The good signal was obtained by utilizing anti-T-T dimer serum (IgG frafction), peroxidase-labeled goat anti-rabbit IgG antibody, and diaminobenzidine-4-HCl and H2O2 as substrates. The anti-sense strand synthetic probe hybridized with cellular RNA of cultured cells with c-yes-1 mRNA, while the sense strand synthetic probe did not. The c-yes-1 mRNA was clearly detected in the cytoplasm of c-yes-1 gene product-expressing cultured cells by in situ hybridization using a T-T dimerized synthetic probe. Synthetic T-T dimerized oligodeoxynucleotides appear to constitute attractive reagents for in situ hybridization studies when supported by appropriate control procedures.
- 日本組織細胞化学会の論文
著者
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Koji Takehiko
Department Of Histology And Cell Biology Nagasaki University Graduate School Of Biomedical Sciences
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MORI Shigeo
Department of Pathology, Institute of Medical Science, University of Tokyo
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Sugawara Isamu
Department of Molecular Pathology The Research Institute of Tuberculosis
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Sugawara I
The Res. Inst. Of Tuberculosis Japan Anti-tuberculosis Assoc.
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Sugawara I
Department Of Pathology Saitama Medical Center Saitama Medical School
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Sugawara Isamu
The Research Institute Of Tuberculosis Japan Anti-tuberculosis Association
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Sugawara Isamu
東京大学医科学研究所
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Sugawara Isamu
Department Of Pathology Institute Of Medical Science University Of Tokyo
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Nakane Paul
長崎大学 解剖3
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Koji Takehiko
Department Of Cell Biology Tokai University School Of Medicine
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UCHINO KENJI
Department of Pathology, The Institute of Medical Science,The University of Tokyo
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SUKEGAWA JUN
Department of Cancer Prevention, The Institute of Medical Science,The University of Tokyo
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NAKANE PAUL
The Third Department of Anatomy, Nagasaki University School of Medicine
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Sugawara Isamu
Department Of Molecular Pathology Reserch Institute Of Tuberculslosis
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Mori Shigeo
Department Of Materials Science Osaka Prefecture University
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Koji Takehiko
Department Fo Histology And Cell Biology Nagasaki University School Of Medicine
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NAKANE Paul
Environmental Biotechnology Institute, California Polytechnic State University
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Sukegawa Jun
Department Of Cancer Prevention The Institute Of Medical Science The University Of Tokyo
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Nakane P
Essential Therapeutics Ca Usa
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Mori Shigeo
Department Of Gastroenterology Shizuoka General Hospital
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Mori Shigeo
Department Of Pathology The Institute Of Medical Science The University Of Tokyo
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Uchino Kenji
Department Of Pathology The Institute Of Medical Science The University Of Tokyo
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Mori Shigeo
Department Of Equilibrium Adaptation Research Division Of Higher Nervous Control Research Institute
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