Initiation of DNA replication in nuclei and purified DNA by a cell-free extract of Xenopus eggs

J. Julian Blow, Ronald A. Laskey

    Research output: Contribution to journalArticle

    415 Citations (Scopus)

    Abstract

    We demonstrate that cell-free extracts prepared from activated eggs of X. laevis by a method similar to that of Lohka and Masui initiate and complete semiconservative DNA replication of sperm nuclei and plasmid DNA. The efficiency of replication is comparable to that in the intact egg. Under optimal conditions 70%-100% of nuclei, and up to 38% of naked DNA molecules replicate completely. Genuine initiation of replication occurs rather than elongation of preformed primers or priming of irreversibly denatured templates. Rereplication of templates is observed under certain conditions. In addition to replicating DNA, these extracts also assemble nucleus-like structures from naked DNA.
    Original languageEnglish
    Pages (from-to)577-587
    Number of pages11
    JournalCell
    Volume47
    Issue number4
    DOIs
    Publication statusPublished - 21 Nov 1986

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    Xenopus
    Cell Extracts
    DNA Replication
    Eggs
    DNA
    Ovum
    Spermatozoa
    Plasmids
    Elongation
    Molecules

    Cite this

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    title = "Initiation of DNA replication in nuclei and purified DNA by a cell-free extract of Xenopus eggs",
    abstract = "We demonstrate that cell-free extracts prepared from activated eggs of X. laevis by a method similar to that of Lohka and Masui initiate and complete semiconservative DNA replication of sperm nuclei and plasmid DNA. The efficiency of replication is comparable to that in the intact egg. Under optimal conditions 70{\%}-100{\%} of nuclei, and up to 38{\%} of naked DNA molecules replicate completely. Genuine initiation of replication occurs rather than elongation of preformed primers or priming of irreversibly denatured templates. Rereplication of templates is observed under certain conditions. In addition to replicating DNA, these extracts also assemble nucleus-like structures from naked DNA.",
    author = "Blow, {J. Julian} and Laskey, {Ronald A.}",
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    Initiation of DNA replication in nuclei and purified DNA by a cell-free extract of Xenopus eggs. / Blow, J. Julian; Laskey, Ronald A.

    In: Cell, Vol. 47, No. 4, 21.11.1986, p. 577-587.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - Initiation of DNA replication in nuclei and purified DNA by a cell-free extract of Xenopus eggs

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    AU - Laskey, Ronald A.

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    N2 - We demonstrate that cell-free extracts prepared from activated eggs of X. laevis by a method similar to that of Lohka and Masui initiate and complete semiconservative DNA replication of sperm nuclei and plasmid DNA. The efficiency of replication is comparable to that in the intact egg. Under optimal conditions 70%-100% of nuclei, and up to 38% of naked DNA molecules replicate completely. Genuine initiation of replication occurs rather than elongation of preformed primers or priming of irreversibly denatured templates. Rereplication of templates is observed under certain conditions. In addition to replicating DNA, these extracts also assemble nucleus-like structures from naked DNA.

    AB - We demonstrate that cell-free extracts prepared from activated eggs of X. laevis by a method similar to that of Lohka and Masui initiate and complete semiconservative DNA replication of sperm nuclei and plasmid DNA. The efficiency of replication is comparable to that in the intact egg. Under optimal conditions 70%-100% of nuclei, and up to 38% of naked DNA molecules replicate completely. Genuine initiation of replication occurs rather than elongation of preformed primers or priming of irreversibly denatured templates. Rereplication of templates is observed under certain conditions. In addition to replicating DNA, these extracts also assemble nucleus-like structures from naked DNA.

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