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Nucleic Acids Research, 1989, Vol. 17, No. 21 8485-8493
© 1989

MOLECULAR BIOLOGY

High level gene expression in mammalian cells by a nuclear T7-phage RNA polymerase

Andre Lieber, Udo Kiessling1 and Michael Strauss

Akademie der Wissenschaften der DDR, Zentralinstitut für Molekularbiologie, Abteilung Molekulare Zellgenetik Berlin-Buch, DDR-1115, GDR 1Abteilung Virologie Berlin-Buch, DDR-1115, GDR

Received August 11, 1989. Revised October 6, 1989. Accepted October 6, 1989.

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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