Overview of Protein Expression in Saccharomyces cerevisiae

Robert L. Strausberg1, Susan L. Strausberg2

1 National Institutes of Health, Bethesda, 2 University of Maryland, Rockville
Publication Name:  Current Protocols in Protein Science
Unit Number:  Unit 5.6
DOI:  10.1002/0471140864.ps0506s02
Online Posting Date:  May, 2001
GO TO THE FULL TEXT: PDF or HTML at Wiley Online Library

Abstract

This overview presents vectors and host strains that are available to direct gene expression in S. cerevisiae, including information on promoters, vector maintenance and copy number, transcription terminators, and selectable markers. Challenges to the expression of foreign proteins are also covered, including attainment of desired production yield, production of protein with appropriate postā€translational modifications, conformation and function, and secretion to the extracellular medium.

     
 
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Table of Contents

  • Vectors and Host Strains
  • Challenges to the Expression of Foreign Proteins
  • Time Frame and Future Prospects
  • Literature Cited
  • Figures
  • Tables
     
 
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Materials

GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Figures

Videos

Literature Cited

Literature Cited
   Ballou, L., Cohen, R., and Ballou, C. 1980. Saccharomyces cerevisiae mutants that make mannoproteins with a truncated carbohydrate outer chain. J. Biol. Chem. 255:5986‐5991.
   Barr, P.J., Steimer, K.S., Sabin, E.A., Parkes, D., George‐Nascimento, C., Stephans, J.C., Powers, M.A., Gyenes, A., Nan Nest, G.A., Miller, E.T., Higgins, K.W., and Luciw, P.A. 1987. Antigenicity and immunogenicity of domains of the human immunodeficiency virus (HIV) envelope protein expressed in the yeast Saccharomyces cerevisiae. Vaccine 5:90‐101.
   Beggs, J.D. 1978. Transformation of yeast by a replicating hybrid plasmid. Nature 275:104‐109.
   Brake, A.J., Merryweather, J.P., Coit, D.G., Heberlein, U.A., Masiarz, F.R., Mullenbach, G.T., Urdea, M.S., Valenzuela, P., and Barr, P.J. 1984. α‐factor‐directed synthesis and secretion of mature foreign proteins in Saccharomyces cerevisiae. Proc. Natl. Acad. Sci. U.S.A. 81:4642‐4646.
   Castelli, L.A., Mardon, C.J., Strike, P.M., Azad, A.A., and Macreadie, I.G. 1994. High‐level secretion of correctly processed beta‐lactamase from Saccharomyces cerevisiae using a high‐copy‐number secretion vector. Gene 142:113‐117.
   Erhart, E. and Hollenberg, C.P. 1983. The presence of a defective LEU2 gene in 2µm DNA recombinant plasmids is responsible for curing and high copy number. J. Bacteriol. 156:625‐635.
   Etcheverry, T. 1990. Induced expression using yeast copper metallothionein promoter. Methods Enzymol. 185:319‐329.
   Haguenauer‐Tsapis, R. and Hinnen, A. 1984. A deletion that includes the signal peptidase cleavage site impairs processing, glycosylation, and secretion of cell‐surface yeast acid phosphatase. Mol. Cell. Biol. 4:2668‐2675.
   Henderson, R.C.A., Cox, B.S., and Tubb, R. 1985. The transformation of brewing yeasts with a plasmid containing the gene for copper resistance. Curr. Genet. 9:133‐138.
   Hinnen, A., Meyhack, B., and Heim, J. 1989. Heterologous gene expression in yeast. In Yeast Genetic Engineering (P.J. Barr, A.J. Brake, and P. Valenzuela, eds.) pp. 193‐213. Butterworths, London.
   Hitzeman, R.A., Chen, C.Y., Dowbenko, D.J., Renz, M.E., Lui, C., Pai, R., Simpson, N.J., Kohr, W.J., Singh, A., Chisholm, V., Hamilton, R., and Chang, C.N. 1990. Use of heterologous and homologous signal sequences for secretion of heterologous proteins from yeast. Methods Enzymol. 185:421‐440.
   Innis, M.A., Holland, M.J., McCabe, P.C., Cole, G.E., Wittman, V.P., Tal, R., Watt, K.W.K., Gelfand, D.H., Holland, J.P., and Meade, J.H. 1985. Expression, glycosylation, and secretion of an Aspergillus glucoamylase by Saccharomyces cerevisiae. Science 228:21‐26.
   Johnston, M. 1987. A model fungal gene regulatory mechanism: The GAL genes of Saccharomyces cerevisiae. Microbiol. Rev. 51:458‐47.
   Kingsman, S.M., Kingsman, A.J., Dobson, M.J., Mellor, J., and Roberts, N.A. 1985. Heterologous gene expression in Saccharomyces cerevisiae. Biotechnol. Genet. Eng. Revs. 3:377‐416.
   Kramer, R.A., DeChiara, T.M., Schaber, M.D., and Hilliker, S. 1984. Regulated expression of a human interferon gene in yeast: Control by phosphate concentration or temperature. Proc. Natl. Acad. Sci. U.S.A. 81:367‐370.
   Kukuruzinska, M.A., Bergh, M.L.E., and Jackson, B.L. 1987. Protein glycosylation in yeast. Annu. Rev. Biochem. 56:915‐944.
   Lemontt, J.F., Wei, C.M., and Dackowski, W.R. 1985. Expression of active human tissue plasminogen activator in yeast. DNA 4:419‐428.
   Lopes, T.S., Klootwijk, J., Veenstra, A.E., van der Aar, P.C., van Heerikhuizen, H., Raue, H.A., and Planta, R.J. 1989. High‐copy‐number integration into the ribosomal DNA of Saccharomyces cerevisiae: A new vector for high‐level expression. Gene 79:199‐206.
   Penttila, M.E., Andr´, L., Lehtovaara, P., Bailey, M., Teeri, T.T., Knowles, J.K.C. 1988. Efficient secretion of two fungal cellobiohydrolases by Saccharomyces cerevisiae. Gene 63:103‐112.
   Robinson, A.S., Hines, V., and Wittrup, K.D. 1994. Protein disulfide isomerase overexpression increases secretion of foreign proteins in Saccharomyces cerevisiae. Bio/Technology 12:381‐384.
   Romanos, M.A., Scorer, C.A., and Clare, J.J. 1992. Foreign gene expression in yeast: A review. Yeast 8:423‐488.
   Sakai, A., Ozawa, F., Higashizaki, T., Shimizu, Y., and Hishinuma, F. 1991. Enhanced secretion of human nerve growth factor from Saccharomyces cerevisiae using an advanced δ‐integration system. Bio/Technology 9:1382‐1385.
   Shuster, J.R. 1989. Regulated transcriptional systems for the production of proteins in yeast: Regulation by carbon source. In Yeast Genetic Engineering (P.J. Barr, A.J. Brake and P. Valenzuela eds.) pp. 83‐108. Butterworths, London.
   Smith, R.A., Duncan, M.J., and Moir, D.T. 1985. Heterologous protein secretion from yeast. Science 230:1219‐1224.
   Strausberg, R.L. and Strausberg, S.L. 1991. Composite yeast vectors. U. S. Patent 5,013,652.
   Tuite, M.F., Dobson, M.J., Roberts, N.A., King, R.M., Burke, D.C., Kingsman, S.M., and Kingsman, A.J. 1982. Regulated high‐efficiency expression of human interferon‐α in Saccharomyces cerevisiae. EMBO J. 1:603‐608.
   Van Arsdell, J.N., Kwok, S., Schweikart, V.L., Ladner, M.B., Gelfand, D.H., Innis, M.A. 1987. Cloning, characterization, and expression in Saccharomyces cerevisiae of endoglucanase I from Trichoderma reesei. Bio/Technology 5:60‐64.
   Wanker, E., Klingsbichel, E., and Schwab, H. 1995. Efficient secretion of Bacillus subtilis levanase by Saccharomyces cerevisiae. Gene 161:45‐49.
   Webster, T.D. and Dickson, R.C. 1983. Direct selection of Saccharomyces cerevisiae resistant to the antibiotic G418 following transformation with a DNA vector carrying the kanamycin resistance gene of Tn 903. Gene 26:243‐252.
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library