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Nucleic Acids Research, 1983, Vol. 11, No. 8 2287-2302
© 1983

MOLECULAR BIOLOGY

Expression in Saccharomyces cerevisiae of human interferon-alpha directed by the TRP1 5' region

M.J. Dobson*, M.F. Tuite, J. Mellor, N.A. Roberts, R.M. King1, D.C. Burke1, A.J. Kingsman and S.M. Kingsman

Department of Biochemistry South Parks Road, Oxford OX1 3QU 1Department of Biological Sciences, University of Warwick Coventry CV4 7AL, UK

*To whom correspondence should be sent

Received February 28, 1983. Accepted March 16, 1983.

The complete 5' flanking region of the yeast TRP1 gene encoding N-(5'-phosphoribosyl)- anthranilate isomerase, a non–abundant protein, has been cloned and the nucleotide sequence data has been extended from –102 to –440. The CT block--CAAG structure common to all efficiently expressed yeast genes is altered in the 5' region of TRP1 and a sequence postulated to be involved in general amino acid regulation is absent. There are two possible TATA boxes at –224 and –;262. TRP1, in common with HIS3, HIS4 and TRP5 has a region of dyad symmetry upstream of the coding sequence which may play a role in initiation of transcription. The relative efficiency of gene expression directed by the complete 5' TRP1 region was assessed by comparison with that of PGK by inserting a cDNA for a human interferon-alpha downstream of their respective 5' regions. The respective interferon yields indicate that their in vivo expression capabilities are a function of their 5' regions.


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