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Plasmid for delivery of nucleic acids to cells and methods of use

  • US 5,763,270 A
  • Filed: 06/07/1995
  • Issued: 06/09/1998
  • Est. Priority Date: 06/07/1995
  • Status: Expired due to Term
First Claim
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1. A plasmid for delivery of a nucleic acid sequence to a eukaryotic cell, comprising:

  • (a) a region which regulates plasmid copy number, wherein said region includes an origin of replication sequence;

    (b) a region which regulates selection of a plasmid containing cell;

    (c) a linker region connecting said region which regulates plasmid copy number to said region which regulates selection of a plasmid containing cell, wherein said linker region includes a site for inserting said nucleic acid sequence, and a transcriptional terminator element to ensure transcription of said region which regulates selection of a plasmid containing cell does not transcribe through said site for insertion of said nucleic acid sequence;

    (d) a plurality of nucleic acid restriction sites flanking said origin of replication of said region which regulates plasmid copy number, said linker region, and said transcriptional terminator sequence; and

    (e) a functional element inserted in a said nucleic acid restriction site, wherein said functional element is selected from the group consisting of a partitioning locus which helps ensure that at least one copy of said plasmid is transmitted to each new cell during cell division and a multimer resolution locus which promotes plasmid monomerization,wherein said region which regulates plasmid copy number, said linker region, and said region which regulates selection of a plasmid containing cell are positioned 5'"'"' to 3'"'"' in the order;

    region which regulates plasmid copy number, linker region, region which regulates selection of a plasmid containing cell, and wherein the transcriptional orientation of said region which regulates selection of a plasmid containing cell is opposite to the transcriptional orientation of a said nucleic acid sequence inserted in said linker region.

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