A New and Efficient Method for Gene Transfer into Mouse FM3A Cells Using Metaphase Chromosomes by Electroporation
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概要
- 論文の詳細を見る
We introduced chromosome-mediated genes into mouse thymidine kinase-deficient FM3A (FM3Atk^-) cells, by electroporation. The effects of some parameters on the electric shock-mediated transfection of FM3Atk^- cells were investigated. Gene transfer of mouse L929 metaphase chromosome DNA into FM3Atk^- resulted in a maximum frequency of (3.0±0.3)×10^<-5> at a cell density of 2.0×10^8/ml and chromosome dosage of 5.0×10^7 cell equivalents/ml in a buffer containing 0.25 M mannitol, 0.5 mM MgCl_2, 0.1 mM CaCl_2, and 1 mM Tris-HCl (pH 7.1). The highest yield of the transformants was obtained at an electric field strength of 1 kV/cm and a capacitance of 35 μF, with a single exponentially decaying pulse at 0℃. The incubation conditions of 60 min at 0℃ was optimal for post-shock incubation after electroporation. The tk gene was detected in the transformants by in situ hybridization analysis.
- 社団法人日本農芸化学会の論文
- 1996-11-23
著者
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Tsuchida Kenji
Memory Lsi Research & Development Center Toshiba Corporation Semiconductor Company
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Tsuchida Kenji
Department Of Applied Physics And Chemistry Faculty Of Engineering Fukui University Of Technology
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Kimoto Hisashi
Department Of Biochemistry I Fukui Medical School
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Kimoto Hisashi
Department Of Applied Physics And Chemistry Fukui Institute Of Technology
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Kimoto Hisashi
Department Of Biochemistry I Faculty Of Medicine Fukui Medical School
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Taketo A
Department Of Applied Physics And Chemistry Fukui University Of Technology
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Taketo Akira
Department Of Applied Physice And Chemistry
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Ohse Morimasa
Department Of Applied Physics And Chemistry Faculty Of Engineering Fukui University Of Technology
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Kusaoke Hideo
Department Of Applied Physics And Chemistry Faculty Of Engineering Fukui University Of Technology
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TOMITA Hideo
Department of Applied Physics and Chemistry, Faculty of Engineering, Fukui University of Technology
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Tomita Hideo
Department Of Applied Physics And Chemistry Faculty Of Engineering Fukui University Of Technology
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