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  • Title: Transcription from plasmid expression vectors is increased up to 14-fold when plasmids are transfected as concatemers.
    Author: Leahy P, Carmichael GG, Rossomando EF.
    Journal: Nucleic Acids Res; 1997 Jan 15; 25(2):449-50. PubMed ID: 9016578.
    Abstract:
    A protocol for increasing transcription from plasmid expression vectors is presented. A vector containing chloramphenicol acetyltransferase (CAT) gene was digested leaving the transcription cassette intact. Heat inactivation of restriction enzymes followed by ligation of the digestion products yielded concatemers which migrated as a single band in agarose gel electrophoresis. Mouse fibroblasts transfected with the concatemers gave a CAT activity that was 14-fold greater than that of cells transfected with a similar mass (equimolar gene number) of the native plasmid. The effect was independent of promoter type, restriction enzyme, number of restriction sites and with a noted exception, cell line.
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