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Nucleic Acids Research, 1984, Vol. 12, No. 17 6797-6812
© 1984

MOLECULAR BIOLOGY

Cloning with tandem gene systems for high level gene expression

Nancy Lee*, J. Cozzitorto, N. Wainwright and D. Testa

Department of Molecular Genetics, Interferon Sciences, Inc. 783 Jersey Avenue, New Brunswick NJ 08901, USA

*To whom reprint requests should be addressed

Received July 27, 1984. Accepted August 13, 1984.

A method has been devised for increasing the copy number of a gene (or genes) cloned into a plasmid while minimizing the size of the plasmid. If n copies of a transcriptional unit are cloned, including the promoter, coding region and terminator, the aize of the plasmid will increase by n times the total size of the unit. However, if we borrow the concept of polycistronic operon and sandwich n structural genes, each with its own ribosome binding-site, between a promoter and a transcription terminator, there will be a space saving equivalent to n–1 promoters and n–1 transcription terminators. We have constructed plasmids in which an E. coli lipoprotein promoter is followed by 1 to 4 human leukocyte interferon genes and a transcription terminator.The applications of this method in genetic engineering are discussed.


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