Saccharomyces cerevisiae expression cloning

Reiser SE, Mitsky TA, Gruys KJ (2000) Characterization and cloning of an (R)-specific trans-2,3-enoylacyl-CoA hydratase from Rhodospirillum rubrum and use of this enzyme for PHA production in Escherichia coli. Appl Microbiol Biotechnol 53 209-218 Robert J, Marchesini S, Delessert S, Poirier Y (2005) Analysis of the beta-oxidation of tians-unsaturated fatty acid in recombinant Saccharomyces cerevisiae expressing a peroxisomal PHA synthase reveals the involvement of a reductase-dependent pathway. Biochim Biophys Acta 1734 169-177... 

Hasslacher, M., Schall, M., Hayn, M. et al. (1996) Molecular cloning of the full-length cDNA of (5)-hydroxynitrile lyase from Hevea brasiliensis. Functional expression in Escherichia coli and Saccharomyces cerevisiae and identification of an active site residue. The Journal of Biological Chemistry, 271, 5884-5891. 

Jarvinen N, Maki M, Rabina J et al (2001) Cloning and expression of Helicobacter pylori GDP-l-fucose synthesizing enzymes (GMD and GMER) in Saccharomyces cerevisiae. Eur J Biochem 268 6458-6464... 

Wamecke, D., Erdmann, R., Fahl, A., Hube, B., Muller, R, Zank, T., Zahringer, U. and Heinz, E. (1999) Cloning and functional expression of UGT genes encoding sterol glucosyltransferases from Saccharomyces cerevisiae, Candida albicans, Pichia pastoris, and Dictyostelium discoideum. J. Biol. Chem., 274,13048-59. 

Saccharomyces cerevisiae is quite commonly used as a host for expression of proteins from cloned genes from yeasts and other organisms. Because flavocytochrome 62 is a yeast enzyme and procedures for isolation of intact enzyme from yeast had already been developed, Reid et al. (143) developed a system for expression of S. cerevisiae flavocy-... 

In the next contribution we learn about hands-on experience and recent improvements with different production systems for biopharmaceuticals at Bayer Health-Care. As previously also published in Nature by Heiner Apeler, Head of Expression, an E. coli host/vector system was originally developed for the efficient production of an interleukin-4 variant, but afterwards it was optimized for the expression of other proteins and even Fab fragments. Process development and optimization of the yeast secretory Saccharomyces cerevisiae for expression of a protease inhibitor will also be presented. The focus, however, is on the use of a recently developed mammalian HKBll (hybrid clone of human kidney and B cells) expression system for recombinant human glycoprotein biopharmaceuticals. HKBll is a favorable cell host for the production of human proteins, because it dehvers biopharmaceuticals that are structurally identical to the natural product. The host/vector system supports the production of gram quantities of proteins in a large-scale transient transfection format as well as the development of stable cell fines. These systems together... 

Amore, R., Kotter, P, Kuster, C., Ciriacy, M., Hollenberg, C. P. (1991). Cloning and Expression in Saccharomyces cerevisiae of the NAD(P)H- dependent xylose reductase- encoding gene (XYLl) from the xylose assimilating yeast Pichia stipitis. Gene, 109, 89-97. 

Moes, C. J., Pretorius, 1. S.. Van Zyl, W. H. (1996). Cloning and expression of the Clostridium thermosulfurogenes D-xylose isomerase gene (xylA) in Saccharomyces cerevisiae. Biotechnology Letters, 1S(2), 269-274. 

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