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. 1997 Jan 15;25(2):449–450. doi: 10.1093/nar/25.2.449

Transcription from plasmid expression vectors is increased up to 14-fold when plasmids are transfected as concatemers.

P Leahy 1, G G Carmichael 1, E F Rossomando 1
PMCID: PMC146435  PMID: 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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Selected References

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