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Efficiency of recombination by Cre transient expression in embryonic stem cells: comparison of various promoters.

Efficiency of recombination by Cre transient expression in embryonic stem cells: comparison of various promoters. Research Abstract Details 

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  • Efficiency of recombination by Cre transient expression in embryonic stem cells: comparison of various promoters. Abstract Text:

    k arakiK Araki,t imaizumiT Imaizumi,k okuyamaK Okuyama,y oikeY Oike,k yamamuraK Yamamura,

    The Cre-loxP recombination system of bacteriophage P1 is frequently utilized in genetic manipulation in embryonic stem (ES) cells. The level of Cre expression is critical to induce loxP site-specific recombination in ES cells. To compare the efficiency of recombination, we constructed four Cre expression vectors driven by different promoters: cytomegarovirus/chicken beta-actin (CAG) promoter, human polypeptide chain elongation factor 1alpha (hEF-1alpha) promoter, mouse phosphoglycerate kinase-1 (mPGK) promoter, and polyoma enhancer/herpes simplex virus thymidine kinase (MC1) promoter. We introduced these Cre expression vectors by electroporation into three ES cell lines carrying a single copy of CAG-loxP-chloramphenicol acetyltransferase (CAT) gene-loxP-beta-galactosidase (beta-gal) gene construct. Since the Cre-mediated recombination leads to excision of the CAT gene, the efficiency of recombination can be monitored as beta-gal expression. No selection system was used in the experiments. The maximum recombination frequency was obtained when the CAG promoter was used, followed by the hEF-1alpha promoter, the mPGK promoter and the MC1 promoter in order. These results indicate that the efficiency of recombination in transient expression system correlates with the promoter activity of Cre expression vector. Thus, it is important to choose the promoter for effective recombination by Cre.

    Efficiency of recombination by Cre transient expression in embryonic stem cells: comparison of various promoters. Publishing Authors By Initials

    k arakiK Araki,t imaizumiT Imaizumi,k okuyamaK Okuyama,y oikeY Oike,k yamamuraK Yamamura,

    For similar proteins: viral proteins research abstracts see: proteins: viral proteins research

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    Efficiency of recombination by Cre transient expression in embryonic stem cells: comparison of various promoters. Journal Published:

    PUBLICATION TYPE: Research Support, Non-U.S. Gov

    Journal: Journal of biochemistry

    VOLUME: 122

    Page Numbers: 977-82

    Journal Abbreviation: J. Biochem.

    ISSN: 0021-924X

    DAY: 19

    MONTH: Nov

    YEAR: 1997

    Efficiency of recombination by Cre transient expression in embryonic stem cells: comparison of various promoters. Information

    Number of References:

    LANGUAGE: eng

    NlmUniqueID: 376600

    Efficiency of recombination by Cre transient expression in embryonic stem cells: comparison of various promoters. Keywords Mesh Terms:

    KEYWORDS: Viral Proteins

    MESH TERMS: genetics

    Chemical & Substance for Abstract: Efficiency of recombination by Cre transient expression in embryonic stem cells: comparison of various promoters. Information

    Substance Name: Integrases

    Registry Number: EC 2.7.7.-

    Grant and Affiliation Information for Efficiency of recombination by Cre transient expression in embryonic stem cells: comparison of various promoters.

    AFFILIATION: Department of Developmental Genetics, Institute of Molecular Embryology and Genetics, Kumamoto University School of Medicine.

    Country: JAPAN

    JAPAN Research PublicationJAPAN Research Publication

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    MEDLINETA: J Biochem

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