Summary
The E. coli aroA gene was inserted between yeast promoter and terminator sequences in different shuttle expression plasmids and found to confer enhanced EPSP synthase activity as well as resistance to glyphosate toxicity. Subsequently, a transformation system using these newly constructed vectors in yeast was characterized. The efficiency of the glyphosate resistance marker for transformation and selection with plasmid pHR6/20-1 in S. cerevisiae laboratory strain SHY2 was found to be relatively high when compared with selection for LEU2 prototrophy. The fate of the recombinant plasmid pHR6/20-1 in the transformants, the preservation of the aroA E. coli DNA fragment in yeast, mitotic stability, EPSP synthase activity, and growth on glyphosate-containing medium have been investigated. As this plasmid also allows direct selection for glyphosate resistant transformants on rich media, the glyphosate resistance marker was used for transforming both S. cerevisiae laboratory strain SHY2 and brewer's yeast strains S. cerevisiae var. “uvarum” BHS5 and BHS2. In all cases, the vector pHR6/20-1 was maintained as an autonomously replicating plasmid. The resistance marker is, therefore, suitable for transforming genetically unlabeled S. cerevisiae laboratory, wild, and industrial yeast strains.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- EPSP :
-
5-enolpyruvylshikimate 3-phosphate
References
Ammerer G (1983) In: Wu e, Grossman L, Moldave K (eds) Methods in enzymology, vol 101c. Academic Press, New York London, pp 192–201
Birnboim HC, Doly J (1979) Nucleic Acids Res 7:1513–1523
Bode R, Kunze G, Birnbaum D (1985) Biochem Physiol Pflanz 180:613–619
Bode R, Schauer F, Birnbaum D (1986) Biochem Physiol Pflanz 181:39–46
Boros I, Lucacsovich T, Baliko G, Venetianer P (1986) Gene 42:97–100
Botstein D, Falco SC, Stewart SE, Brennan M, Scherer S, Stinchcomb DT, Struhl K, Davis RW (1979) Gene 8:17–24
Case ME (1983) In: Lurquin PF, Leinhofs A (eds) Genetic engineering in eukaryotes. Plenum Press, New York, pp 1–5
Duncan K, Coggins JR (1984) Biochem Soc Trans 12:274–275
Duncan K, Coggins JR (1986) Biochem J 234:49–57
Duncan K, Lewendon A, Coggins JR (1984) FEBS Lett 165:121–127
Fogel S, Welch JW, Cathala G, Karin M (1983) Curr Genet 7:347–355
Gatignol A, Baron M, Tiraby G (1987) Mol Gen Genet 207:342–348
Gollub E, Zalkin H, Sprinson DB (1967) J Biol Chem 242:5323–5333
Gritz L, Davis J (1983) Gene 25:179–188
Hadfield C, Cashmore AM, Meacock PA (1987) Gene 52:59–70
Hanahan D (1983) J Mol Biol 166:557–580
Hinnen A, Hicks JB, Fink GR (1978) Proc Natl Acad Sci USA 75:1929–1933
Hitzeman RA, Clarke L, Carbon J (1980) J Biol Chem 255:12073–12080
Hollaender-Czytko H, Johanning D, Meyer HE, Amrhein N (1988) Plant Mol Biol 11:215–220
Ito H, Fukada Y, Murata K, Kimura A (1983) J Bacteriol 153:163–168
Jaworski EG (1972) J Agric Food Chem 20:1195–1198
Jiminez A, Davis J (1980) Nature 287:869–871
Kaster KR, Burgett SG, Ingolia TD (1984) Curr Genet 8:353–358
Knowles PF, Sprinson DB (1970) In: Tabor H, Tabor GW (eds) Methods in enzymology, vol 17A. Academic Press, New York London, pp 351–352
Kunze G, Hecker M, Birnbaum D (1984) Z Allg Mikrobiol 24:33–40
Larimer FW, Morse CC, Beck AK, Cole KW, Gaertner FH (1983) Mol Cell Biol 3:1609–1614
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) J Mol Chem 193:265–275
Struhl K, Stinchcomb DT, Scherer S, Davis RW (1979) Proc Natl Acad Sci USA 76:1035–1039
Zhu J, Contreras R, Gheysen D, Erst J, Fiers W (1985) Biotechnology 3:451–456
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Kunze, G., Bode, R., Rintala, H. et al. Heterologous gene expression of the glyphosate resistance marker and its application in yeast transformation. Curr Genet 15, 91–98 (1989). https://doi.org/10.1007/BF00435454
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00435454