title: Expression of soluble active human beta 1,4 galactosyltransferase in Saccharomyces cerevisiae.
abstract: Sequences coding for the cytoplasmic and transmembrane domains were removed 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 fused to sequences coding for the yeast invertase signal sequence. The hybrid was inserted together with a constitutive yeast promoter and a terminator into a E. coli/yeast shuttle vector. Saccharomyces cerevisiae strain BT150 transformed with this new expression vector expressed enzymically active soluble enzyme, whereas no activity was detectable in mock-transformed yeasts. The enzyme product was identified by HPLC analysis and shown to correspond to the expected product N-acetyllactosamine.
authors.affiliation: Physiologisches Institut, Universität Zürich, Switzerland.
authors.forename: R
authors.initials: R
authors.lastname: Kleene
authors.forename: C H
authors.initials: CH
authors.lastname: Krezdorn
authors.forename: G
authors.initials: G
authors.lastname: Watzele
authors.forename: B
authors.initials: B
authors.lastname: Meyhack
authors.forename: G F
authors.initials: GF
authors.lastname: Herrmann
authors.forename: C
authors.initials: C
authors.lastname: Wandrey
authors.forename: E G
authors.initials: EG
authors.lastname: Berger
mesh_terms: D001483:Base Sequence; D003001:Cloning, Molecular; D005700:Galactosyltransferases / Q000737:chemistry / Q000235:genetics*; D015870:Gene Expression; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D012333:RNA, Messenger / Q000235:genetics; D011994:Recombinant Proteins; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D012995:Solubility; D013329:Structure-Activity Relationship
chemical_list: D009838:Oligodeoxyribonucleotides; D012333:RNA, Messenger; D011994:Recombinant Proteins; D005700:Galactosyltransferases