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Efficient intra- and extracellular production of human beta-1,4-galactosyltransferase in Saccharomyces cerevisiae is mediated by yeast secretion leaders. 1994

T Schwientek, and J F Ernst
Institut für Mikrobiologie, Heinrich-Heine-Universität, Düsseldorf, Germany.

We have compared the function of homologous and heterologous secretion leaders to mediate production of human beta-1,4-galactosyltransferase (Gal-Tf) in the yeast Saccharomyces cerevisiae. Although all genes encoding leader/Gal-Tf fusions were transcribed by strong yeast promoters, only low production levels were obtained for full-length Gal-Tf containing the human membrane-anchor region. In contrast, a gene fusion encoding the membrane-anchor region of the yeast alpha-1,2-mannosyltransferase (Mnt1) fused to soluble Gal-Tf yielded high mRNA and intracellular protein levels. Gal-Tf could also be produced extracellularly using a fusion of the pre-pro region of the yeast Mf alpha 1 precursor (MF alpha 1) to soluble Gal-Tf; a fusion containing only the pre-region of Mf alpha 1 was synthesized intracellularly, but did not lead to Gal-Tf activity in the culture medium. All yeast-produced Gal-Tf proteins were enzymatically active. These results demonstrate that yeast secretion leaders are advantageous to achieve efficient production of active Gal-Tf in S. cerevisiae.

UI MeSH Term Description Entries
D008364 Mannosyltransferases Enzymes that catalyze the transfer of mannose from a nucleoside diphosphate mannose to an acceptor molecule which is frequently another carbohydrate. The group includes EC 2.4.1.32, EC 2.4.1.48, EC 2.4.1.54, and EC 2.4.1.57. Mannosyltransferase
D009237 N-Acetyllactosamine Synthase The A protein of the lactose synthase complex. In the presence of the B protein (LACTALBUMIN) specificity is changed from N-acetylglucosamine to glucose. EC 2.4.1.90. N-Acetyllactosamine Synthetase,UDP Galactose Acetylglucosamine Galactosyltransferase,N-Acetylglucosamine beta-D-Galactosyltransferase,UDP Galactose-N-Acetylglucosamine Galactosyltransferase,beta-1,4-Galactosyltransferase,Galactosyltransferase, UDP Galactose-N-Acetylglucosamine,N Acetylglucosamine beta D Galactosyltransferase,N Acetyllactosamine Synthase,N Acetyllactosamine Synthetase,Synthase, N-Acetyllactosamine,Synthetase, N-Acetyllactosamine,UDP Galactose N Acetylglucosamine Galactosyltransferase,beta 1,4 Galactosyltransferase,beta-D-Galactosyltransferase, N-Acetylglucosamine
D011993 Recombinant Fusion Proteins Recombinant proteins produced by the GENETIC TRANSLATION of fused genes formed by the combination of NUCLEIC ACID REGULATORY SEQUENCES of one or more genes with the protein coding sequences of one or more genes. Fusion Proteins, Recombinant,Recombinant Chimeric Protein,Recombinant Fusion Protein,Recombinant Hybrid Protein,Chimeric Proteins, Recombinant,Hybrid Proteins, Recombinant,Recombinant Chimeric Proteins,Recombinant Hybrid Proteins,Chimeric Protein, Recombinant,Fusion Protein, Recombinant,Hybrid Protein, Recombinant,Protein, Recombinant Chimeric,Protein, Recombinant Fusion,Protein, Recombinant Hybrid,Proteins, Recombinant Chimeric,Proteins, Recombinant Fusion,Proteins, Recombinant Hybrid
D002451 Cell Compartmentation A partitioning within cells due to the selectively permeable membranes which enclose each of the separate parts, e.g., mitochondria, lysosomes, etc. Cell Compartmentations,Compartmentation, Cell,Compartmentations, Cell
D005822 Genetic Vectors DNA molecules capable of autonomous replication within a host cell and into which other DNA sequences can be inserted and thus amplified. Many are derived from PLASMIDS; BACTERIOPHAGES; or VIRUSES. They are used for transporting foreign genes into recipient cells. Genetic vectors possess a functional replicator site and contain GENETIC MARKERS to facilitate their selective recognition. Cloning Vectors,Shuttle Vectors,Vectors, Genetic,Cloning Vector,Genetic Vector,Shuttle Vector,Vector, Cloning,Vector, Genetic,Vector, Shuttle,Vectors, Cloning,Vectors, Shuttle
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D001692 Biological Transport The movement of materials (including biochemical substances and drugs) through a biological system at the cellular level. The transport can be across cell membranes and epithelial layers. It also can occur within intracellular compartments and extracellular compartments. Transport, Biological,Biologic Transport,Transport, Biologic
D012441 Saccharomyces cerevisiae A species of the genus SACCHAROMYCES, family Saccharomycetaceae, order Saccharomycetales, known as "baker's" or "brewer's" yeast. The dried form is used as a dietary supplement. Baker's Yeast,Brewer's Yeast,Candida robusta,S. cerevisiae,Saccharomyces capensis,Saccharomyces italicus,Saccharomyces oviformis,Saccharomyces uvarum var. melibiosus,Yeast, Baker's,Yeast, Brewer's,Baker Yeast,S cerevisiae,Baker's Yeasts,Yeast, Baker
D015152 Blotting, Northern Detection of RNA that has been electrophoretically separated and immobilized by blotting on nitrocellulose or other type of paper or nylon membrane followed by hybridization with labeled NUCLEIC ACID PROBES. Northern Blotting,Blot, Northern,Northern Blot,Blots, Northern,Blottings, Northern,Northern Blots,Northern Blottings
D021382 Protein Sorting Signals Amino acid sequences found in transported proteins that selectively guide the distribution of the proteins to specific cellular compartments. Leader Signal Peptides,Leader Peptide,Leader Sequences, Peptide,Peptide Leader Sequences,Peptide Signal Sequences,Signal Peptide,Signal Peptides,Signal Sequence, Peptide,Signal Sequences,Signal Sequences, Peptide,Leader Peptides,Leader Sequence, Peptide,Leader Signal Peptide,Peptide Leader Sequence,Peptide Signal Sequence,Peptide, Leader,Peptide, Leader Signal,Peptide, Signal,Peptides, Leader,Peptides, Leader Signal,Peptides, Signal,Protein Sorting Signal,Sequence, Peptide Leader,Sequence, Peptide Signal,Sequence, Signal,Sequences, Peptide Leader,Sequences, Peptide Signal,Sequences, Signal,Signal Peptide, Leader,Signal Peptides, Leader,Signal Sequence,Signal, Protein Sorting,Signals, Protein Sorting,Sorting Signal, Protein,Sorting Signals, Protein

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