Skip to main content
SpringerLink
Log in

Cloning and expression on a multicopy vector of five invertase genes of Saccharomyces cerevisiae

  • Original Articles
  • Published:
Current Genetics Aims and scope Submit manuscript

Summary

Six unlinked loci for invertase structural genes are known in the yeast Saccharomyces cerevisiae: SUC1-SUC5 and SUC7. These genes are similar in structure and expression but not identical. Different yeast strains possess none, one or several of these genes.

We have isolated the genes SUC1-SUC5, subcloned them into the multicopy vector YEp24 and compared the expression of the five SUC genes in one recipient strain.

SUC2 was isolated by transformation of a suc0 strain with a gene pool and complementation to sucrose fermentation. SUC4 was cloned from a minipool of chromosomal fragments which were shown to contain SUC4 by Southern hybridization. SUC1, SUC3 and SUC5 were isolated using the method of plasmid eviction. A plasmid containing regions flanking SUC4 was integrated next to these SUC genes. The plasmid together with the SUC genes were then cut out of the chromosome using an appropriate restriction endonuclease.

The length of chromosomal DNA fragments containing the different SUC genes were 4.8 kb for SUC1, 5.2 kb for SUC2, 4.8 kb for SUC3, 12.8 kb for SUC4 and 17.2 kb for SUC5.

Fragments containing the complete SUC genes and the sequences controlling their expression were subcloned into YEp24 and transformed into a strain without any active invertase gene. Invertase activity of transformants was measured after growth repressing (8% glucose) and derepressing (2% raffinose) conditions. As expected from results with strains carrying the individual SUC genes in a chromosomal location, the SUC genes were expressed to a different extent.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Beggs JD (1978) Nature 275:104–108

    Google Scholar 

  • Birnboim HC (1983) Methods Enzymol 100:243–255

    Google Scholar 

  • Bolivar F, Backman K (1979) Methods Enzymol 68:245–267

    Google Scholar 

  • Botstein D, Davis RW (1982) The molecular biology of the yeast Saccharomyces. Cold Spring Harbor Lab, pp 607–636

  • Broach JR, Strathern JN, Hicks JB (1979) Gene 8:121–133

    Google Scholar 

  • Carlson M, Osmond BC, Botstein D (1981a) Genetics 98:25–40

    Google Scholar 

  • Carlson M, Osmond BC, Botstein D (1981b) Genetics 98:41–54

    Google Scholar 

  • Carlson M, Botstein D (1982) Cell 28:145–154

    Google Scholar 

  • Carlson M, Botstein D (1983) Mol Cell Biol 3:351–359

    Google Scholar 

  • Carlson M Celenza J, Eng FJ (1985) Mol Cell Biol 5:2894–2902

    Google Scholar 

  • Clewell DB (1972) J Bacteriol 110:667–679

    Google Scholar 

  • Cohen SN, Chang ACY, Hsu L (1972) Proc Natl Acad Sci USA 69:2110–2114

    Google Scholar 

  • Futcher AB, Cox BS (1984) J Bacteriol 157:283–290

    Google Scholar 

  • Gascón S, Ottolenghi P (1967) CR Trav Lab Carlsberg 36:85–93

    Google Scholar 

  • Gáscon S, Lampen JO (1968) J Biol Chem 243:1567–1572

    Google Scholar 

  • Goldstein A Lampen JO (1975) Methods Enzymol 42:504–511

    Google Scholar 

  • Grossmann MK (1979) Thesis phD TH Darmstadt

  • Grossmann MK, Zimmermann FK (1979) Mol Gen Genet 175:223–229

    Google Scholar 

  • Kaiser CA, Botstein D (1986) Mol Cell Biol 6:2382–2391

    Google Scholar 

  • Lehle L, Cohen RE, Ballou CE (1979) J Biol Chem 254:12209–12218

    Google Scholar 

  • Mortimer R, Hawthorne D (1966) Genetics 53:165–173

    Google Scholar 

  • Nasmyth KA, Reed SI (1980) Proc Natl Acad Sci USA 77:2119–2123

    Google Scholar 

  • Nasmyth KA, Tatchell K (1980) Cell 19:753–764

    Google Scholar 

  • Ottolenghi P (1971) CR Trav Lab Carlsberg 38:213–221

    Google Scholar 

  • Parra F, Herrero P, Moreno F, Gascon S (1980) FEBS Lett 118:330–332

    Google Scholar 

  • Rigby PWJ, Dieckmann M, Rhodes C, Berg P (1977) J Mol Biol 113:237–251

    Google Scholar 

  • Rodriguez L, Lampen JO, MacKay VL (1981) Mol Cell Biol 1:469–474

    Google Scholar 

  • Sarokin L, Carlson M (1984) Mol Cell Biol 4:2750–2757

    Google Scholar 

  • Sarokin L, Carlson M (1985) Nucleic Acid Res 13:6089–6113

    Google Scholar 

  • Struhl K, Stinchcomb DT, Scherer S, Davis RW (1979) Proc Natl Acad Sci USA 76:1035–1039

    Google Scholar 

  • Taussig R, Carlson M (1983) Nucleic Acid Res 11:1943–1954

    Google Scholar 

  • Tautz D, Renz M (1983) Anal Biochem 132:14–19

    Google Scholar 

  • Wahl GM, Stern M, Stark GR (1979) Proc Natl Acad Sci USA 76:3683–3687

    Google Scholar 

  • Winge O, Roberts C (1952) CR Trav Lab Carlsberg 25:141–171

    Google Scholar 

  • Winston F, Chumley F, Fink GR (1983) Methods Enzymol 101:211–227

    Google Scholar 

  • Zamenhoff S (1957) Methods Enzymol 3:696–704

    Google Scholar 

  • Zimmermann FK (1975) Mutat Res 31:71–86

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institut für Mikrobiologie, Technische Hochschule Darmstadt, Schnittspahnstrasse 10, D-6100, Darmstadt, Germany

    Stefan Hohmann & Friedrich K. Zimmermann

Authors
  1. Stefan Hohmann
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Friedrich K. Zimmermann
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Dedicated to Prof. Dr. Fritz Kaudewitz on the occasion of his 65th birthday

This work was supported by Deutsche Forschungsgemeinschaft

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hohmann, S., Zimmermann, F.K. Cloning and expression on a multicopy vector of five invertase genes of Saccharomyces cerevisiae . Curr Genet 11, 217–225 (1986). https://doi.org/10.1007/BF00420610

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00420610

Key words

Search

Navigation