Gene replacement of the p53 gene with the lacz gene in mouse embryonic stem-cells and mice by using two steps of homologous recombination

Yoichi Gondo, Kazuhiko Nakamura, Kazuki Nakao, Toshikuni Sasaoka, Kenichi Ito, Makoto Kimura, Motoya Katsuki

研究成果: ジャーナルへの寄稿記事

67 引用 (Scopus)

抄録

Two steps of gene targeting were used to replace the p53 gene with the E.coli β-galactosidase (lacZ) gene in mouse embryonic stem (ES) cells. The first targeting vector consisted of neo and herpes simplex virus thymidine kinase (HSV-tk) genes as a neo-tk cassette in the middle of the targeting vector. At the first targeting, the homologous recombinants became G418 resistant and ganciclovir (GANC) sensitive and were selected by G418 alone. At the second targeting, homologous recombination reciprocally exchanged the neo-tk cassette in the ES cell chromosome with the lacZ fragment in the second targeting vector and thus made the ES cells GANC resistant. We obtained two ES cell clones, in which the p53 gene for both had been replaced with a totally non-homologous sequence of the lacZ gene. The germ-line transmission of the manipulated ES cells also demonstrated that the entire procedure had no detrimental effects on ES cells at all.

元の言語英語
ページ(範囲)830-837
ページ数8
ジャーナルBiochemical and Biophysical Research Communications
202
発行部数2
DOI
出版物ステータス出版済み - 7 29 1994

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Lac Operon
Homologous Recombination
p53 Genes
Embryonic Stem Cells
Stem cells
Genes
Ganciclovir
Galactosidases
Gene Targeting
Thymidine Kinase
Simplexvirus
Germ Cells
Chromosomes
Viruses
Escherichia coli
Clone Cells
Mouse Embryonic Stem Cells
Electric lines

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

これを引用

Gene replacement of the p53 gene with the lacz gene in mouse embryonic stem-cells and mice by using two steps of homologous recombination. / Gondo, Yoichi; Nakamura, Kazuhiko; Nakao, Kazuki; Sasaoka, Toshikuni; Ito, Kenichi; Kimura, Makoto; Katsuki, Motoya.

:: Biochemical and Biophysical Research Communications, 巻 202, 番号 2, 29.07.1994, p. 830-837.

研究成果: ジャーナルへの寄稿記事

Gondo, Yoichi ; Nakamura, Kazuhiko ; Nakao, Kazuki ; Sasaoka, Toshikuni ; Ito, Kenichi ; Kimura, Makoto ; Katsuki, Motoya. / Gene replacement of the p53 gene with the lacz gene in mouse embryonic stem-cells and mice by using two steps of homologous recombination. :: Biochemical and Biophysical Research Communications. 1994 ; 巻 202, 番号 2. pp. 830-837.
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abstract = "Two steps of gene targeting were used to replace the p53 gene with the E.coli β-galactosidase (lacZ) gene in mouse embryonic stem (ES) cells. The first targeting vector consisted of neo and herpes simplex virus thymidine kinase (HSV-tk) genes as a neo-tk cassette in the middle of the targeting vector. At the first targeting, the homologous recombinants became G418 resistant and ganciclovir (GANC) sensitive and were selected by G418 alone. At the second targeting, homologous recombination reciprocally exchanged the neo-tk cassette in the ES cell chromosome with the lacZ fragment in the second targeting vector and thus made the ES cells GANC resistant. We obtained two ES cell clones, in which the p53 gene for both had been replaced with a totally non-homologous sequence of the lacZ gene. The germ-line transmission of the manipulated ES cells also demonstrated that the entire procedure had no detrimental effects on ES cells at all.",
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AU - Ito, Kenichi

AU - Kimura, Makoto

AU - Katsuki, Motoya

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