The promoter of the beta-glucosidase gene from Kluyveromyces fragilis contains sequences that act as upstream repressing sequences in Saccharomyces cerevisiae.
The relationship between the promoter length of the Kluyveromyces fragilis beta-glucosidase gene and the level of its expression in Saccharomyces cerevisiae was studied by gene fusion between deleted promoter fragments of various lengths and the promoterless beta-galactosidase gene of Escherichia coli. The removal of a region from position -425 to -232 led to a tenfold increase in the expression of the gene. The same results were obtained for the reconstructed beta-glucosidase gene with the same promoter length. It is likely that the deletion of this part of the promoter removes negative regulatory elements which are functional in Saccharomyces cerevisiae. This increase in activity is the main event which may explain the high increase in gene expression (60-fold) previously observed for an upstream deletion obtained during subcloning experiments of the beta-glucosidase gene. It is also shown that the expression of the gene greatly depends upon the nature of the recipient strain, the growth phase of the cell and that of the vector carrying it.
Iborra
F
F
Laboratoire de Biologie et Génétique Moléculaire, Université Paris-Sud, Orsay, France.
Raynal
A
A
Guerineau
M
M
D001483:Base Sequence; D003001:Cloning, Molecular; D005796:Genes; D005800:Genes, Fungal; D005959:Glucosidases / Q000235:genetics*; D007716:Kluyveromyces / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D008969:Molecular Sequence Data; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D004718:Saccharomycetales / Q000235:genetics*; D014158:Transcription, Genetic; D001617:beta-Glucosidase / Q000235:genetics*
D012333:RNA, Messenger; D005959:Glucosidases; D001617:beta-Glucosidase