High level gene expression in mammalian cells by a nuclear T7-phage RNA polymerase (original) (raw)
Journal Article
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Abteilung Virologie
Berlin-Buch, DDR-1115, GDR
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Revision received:
06 October 1989
Accepted:
06 October 1989
Published:
11 November 1989
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Andre Lieber, Udo Kiessling, Michael Strauss, High level gene expression in mammalian cells by a nuclear T7-phage RNA polymerase, Nucleic Acids Research, Volume 17, Issue 21, 11 November 1989, Pages 8485–8493, https://doi.org/10.1093/nar/17.21.8485
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Abstract
Here we describe a novel expression system for mammalian cells which is based on transcription of hybrid genes containing T7 phage promoters by a T7 phage RNA polymerase targeted to the nucleus of the host cells. The RNA polymerase gene of T7 phage has been modified by substituting a sequence encoding the nuclear location signal of SV40 large T antigen for the N-terminal part of the polymerase gene. Expression of the modified gene is driven by the mouse metallothionein promoter in transfected mouse Ltk − cells resulting in high concentration of the polymerase in the nucleus. Nuclear T7 RNA polymerase directs efficient transcription of the cat gene under control of a T7 promoter. T7 constructs are expressed at a level at least 6 fold higher than the prototype pRSVcat. The unique properties of this heterologeous expression system are discussed.
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