Translation of Foreign genes in S. cerevisiae

Brett Burkholder burkhold at gmm.gen.emory.edu
Mon Dec 16 11:11:41 EST 1996


I am having trouble expressing two proteins using a Gal 1/10 promoter in
a gal80 knockout strain.  I'm checking all the obvious problems, such as
gene conversion of host strain with marker gene and promoter efficacy.
(The sequences are unlikely to be toxic to the yeast cells).

I want to know if there are minimum requirements specific to the
translation of cloned genes in yeast:

1). Is there a minimum or maximum length of a 5' UTR necessary for
translation in yeast ?

2) Are there any consensus sequences for context of the initiator codon
in yeast, such as the Kozak sequence for higher eukaryotes ?
(In Marilyn Kozak's review of the "Scanning Model" of translation,
J Cell Bio 108:229-241, 1989, she states that the consensus for higher
eukaryotes seems to have little or no bearing on translation in yeast)

3). Are 3' UTR's necessary for translation, specifically polyadenylation
signals ?

4) Other requirements besides an ORF and above factors ?

Any and all help (references, etc.) will be appreciated.


Brett Burkholder
Emory University
Genetics and Molecular Medicine



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