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Nucleic Acids Research, 1983, Vol. 11, No. 6 1819-1837
© 1983

MOLECULAR BIOLOGY

Expression of the human interferon–{gamma} cDNA in yeast

Rik Derynck, Arjun Singh and David V. Goeddel

Department of Molecular Biology, Genentech, Inc., 460 Point San Bruno Boulevard, South San Francisco, CA 94080, USA.

Received December 8, 1982. Accepted February 22, 1983.

Several plasmids which direct the expression of human interferon–{gamma} (IFN–{gamma}) cDNA in the yeast Saccharomyces cerevisiae under the transcriptional control of the 3–phosphoglycerate kinase (PGK) promoter have been constructed. Up to 25x106 units per liter culture of biologically active IFN-y are synthesized in yeast transformed with these expression plasmids. The amount of IFN–{gamma} mRNA in yeast transformed with the IFN–{gamma} expression plasmids is much lower than the amount of PGK mRNA present when the PGK gene and promoter are inserted on a similar plasmid. The major and three minor sites of transcription initiation have been localized. The major starting point of the IFN–{gamma} mRNA is at 4 basepairs upstream of the PGK mRNA start. All transcription starts map at an ApG as does PGK mRNA. The yeast IFN–{gamma} mRNA terminates in a specific segment of the 3’ untranslated region of the IFN–{gamma} cDNA. The polyadenylation site at about 182-184 nucleotides 3' of the IFN–{gamma} stop codon is preceded by a sequence which is very similar to a sequence proposed to be involved in transcription termination and polyadenylation (49).


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