Bioluminescent PCR-RFLP Enzyme-Linked Immunosorbent Assay for Analysis of Vitamin D Receptor Gene Polymorphism.
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- ARAKAWA Hidetoshi
- School of Pharmaceutical Sciences, Showa University
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- KOKADO Amane
- School of Pharmaceutical Sciences, Showa University
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- YOSHIZAWA Shuzo
- School of Pharmaceutical Sciences, Showa University
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- MAEDA Masako
- School of Pharmaceutical Sciences, Showa University
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- TOKITA Akifumi
- Department of Pediatrics, Juntendo University School of Medicine
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- YAMASHIRO Yuichiro
- Department of Pediatrics, Juntendo University School of Medicine
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説明
We developed a sensitive and rapid PCR-RFLP ELISA using acetate kinase (AK) and firefly luciferase as a detection system. AK used as a label enzyme could sensitively be detected by bioluminescent assay using the firefly luciferase reac-tion. The detection limit was 10-20 mol/assay and the luminescence was stable for 48 h. FITC-labeled sense primer and biotin labeled anti sense primer were used for PCR amplification of the vitamin D receptor gene. After PCR, the products were digested with Taq I or Apa I enzyme. The reaction products were diluted with assay buffer and transferred to a plate coated with anti FITC IgG. After incubation for 2 h at 37°C, the plate was washed and reacted with avidin/biotinylated AK, the AK activity was detected by bioluminescence assay using the firefly luciferin/luciferase system. DNA polymorphism types (AA, Aa, aa, TT, Tt, tt) of the vitamin D receptor gene (VDR) could be clearly determined by measuring the bioluminescent intensity or by using photon imaging with a CCD camera.
収録刊行物
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- Analytical Sciences
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Analytical Sciences 15 (10), 943-949, 1999
社団法人 日本分析化学会
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詳細情報 詳細情報について
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- CRID
- 1390282679233000064
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- NII論文ID
- 10004707766
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- NII書誌ID
- AA10500785
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- COI
- 1:CAS:528:DyaK1MXms1agtrw%3D
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- ISSN
- 13482246
- 09106340
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- NDL書誌ID
- 4872789
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- Web Site
- http://id.ndl.go.jp/bib/4872789
- https://ndlsearch.ndl.go.jp/books/R000000004-I4872789
- https://link.springer.com/content/pdf/10.2116/analsci.15.943.pdf
- https://link.springer.com/article/10.2116/analsci.15.943/fulltext.html
- http://www.jstage.jst.go.jp/article/analsci/15/10/15_10_943/_pdf
- https://search.jamas.or.jp/link/ui/2000117874
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- 本文言語コード
- en
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- データソース種別
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- JaLC
- NDL
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