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A New Vector for Cloning Large Eukaryotic DNA Segments in Escherichia coli

Bio/Technology volume 8, pages 841–844 (1990)Cite this article

Abstract

We investigated the relationship between plasmid size and electroporation efficiency in E. coli and found that even very large plasmids can be transfected efficiently. The efficiencies are well above the minimum required to construct representative libraries of complex eukaryotic genomes. To exploit this observation we constructed a novel mammalian-E. coli shuttle vector whose replication in E. coli is driven by the F sex factor episome origin. This new vector system should accept inserts well in excess of 100 kb, thus putting the cloning of mammalian genes by direct phenotypic complementation within reach.

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Author notes

  1. Eduardo D. Leonardo

    Present address: University of California Medical School, San Francisco, CA

  2. John M. Sedivy: Corresponding author.

Authors and Affiliations

  1. Department of Molecular Biophysics and Biochemistry, Yale University, 333 Cedar Street, New Haven, CT, 06510

    John M. Sedivy

Authors
  1. Eduardo D. Leonardo
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  2. John M. Sedivy
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Leonardo, E., Sedivy, J. A New Vector for Cloning Large Eukaryotic DNA Segments in Escherichia coli. Nat Biotechnol 8, 841–844 (1990). https://doi.org/10.1038/nbt0990-841

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