R. Kleene et al., EXPRESSION OF SOLUBLE ACTIVE HUMAN BETA-1,4 GALACTOSYLTRANSFERASE IN SACCHAROMYCES-CEREVISIAE, Biochemical and biophysical research communications, 201(1), 1994, pp. 160-167
Sequences coding for the cytoplasmic and transmembrane domains were re
moved from the cDNA of the human Golgi resident membrane protein beta
1,4 galactosyltransferase (gal-T). The remaining sequences coding for
the stem and catalytical domains of this glycosyltransferase were fuse
d to sequences coding for the yeast invertase signal sequence. The hyb
rid was inserted together with a constitutive yeast promoter and a ter
minator into a E. coli/yeast shuttle vector. Saccharomyces cerevisiae
strain BT150 transformed with this new expression vector expressed enz
ymically active soluble enzyme, whereas no activity was detectable in
mock-transformed yeasts. The enzyme product was identified by HPLC ana
lysis and shown to correspond to the expected product N-acetyllactosam
ine. (C) 1994 Academic Press, Inc.