Skip to main content
Log in

A hybrid DNA sequence containing the replication origin of the multicopy yeast plasmid 2 μm circle and an additional repeated sequence can convert maltose-negative into maltose-positive strains

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

Yeast DNA pools were prepared by ligating partial Sau3A genomic digests from strains carrying various MAL genes into the BamHI site of the yeast-Escherichia coli shuttle vector YRp7. They were used to transform recipient yeast strains that could not utilize maltose since they lacked a classical MAL gene. Transformants were obtained that could use maltose and also formed normal levels of maltase. They were unstable. They would lose the selective marker TRP1 of YRp7 alone, together with the ability to utilize maltose or only the ability to utilize maltose. The insertion of one of the plasmids was used as a hybridization probe for the others and found to share homologous sequences with all. They were then shown to contain the replication origin of the yeast 2 μm circle plasmid and additional genomic digests of total yeast DNA. They hybridized at various degrees of efficiency with several bands, indicating that they were part of a family of repeated sequences. Apparently, it was the combination of the replication origin of the 2 μm circles with the additional sequences that promoted maltose utilization.

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

Access this article

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

  • Barnett JA (1976) The utilization of sugars by yeasts. Adv Carbohydr Chem Biochem 32:126–234

    Google Scholar 

  • Beggs JD (1978) Transformation of yeast by a replicating hybrid plasmid. Nature 275:104–108

    Google Scholar 

  • Bolivar F, Backman K (1979) Plasmids of Escherichia coli as cloning vectors. Methods Enzymol 68:245–267

    Google Scholar 

  • Broach JR, Strathern JN, Hicks JB (1979) Transformation in yeast: Development of a hybrid cloning vector and isolation of the CAN1 gene. Gene 8:121–133

    Google Scholar 

  • Broach JR (1981) The yeast plasmid 2 μ circle. In: Strathern JN, Jones EW, Broach JR, The molecular biology of the yeast Saccharomyces. Life cycle and inheritance. Cold Spring Harbor Laboratory: 445–470

  • Chow TJ, Goldenthal MJ, Cohen JD, Hedge M, Marmur J (1983) Identification and physical characterization of yeast maltase structural genes. Mol Gen Genet 191:366–371

    Google Scholar 

  • Ciriacy M (1975) Genetics of alcohol dehydrogenase in Saccharomyces cerevisiae. I. Isolation and genetic analysis of adh-mutants. Mut Res 29:315–326

    Google Scholar 

  • Ciriacy M, Breitenbach I (1979) Physiological effects of seven different blocks in glycolysis in Saccharomyces cerevisiae. J Bacteriol 139:152–160

    Google Scholar 

  • Ciriacy M, Williamson VM (1981) Analysis of mutations affecting Ty-mediated gene expression in Saccharomyces cerevisiae. Mol Gen Genet 182:159–163

    Google Scholar 

  • Clewell DB (1972) Nature of Col E1 plasmid replication in Escherichia coli in the presence of chloramphenicol. J Bacteriol 110:667–676

    Google Scholar 

  • Cohen SN, Chang ACY, Hsu L (1972) Nonchromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA. Proc Natl Acad Sci USA 69:2110–2114

    Google Scholar 

  • Denis CL, Ciriacy M, Young ET (1981) A positive regulatory gene is required for accumulation of the functional messenger RNA for glucose-repressible alcohol dehydrogenase from Saccharomyces cerevisiae. J Mol Biol 148:355–368

    Google Scholar 

  • Falco SC, Rose M, Botstein D (1983) Homolgous recombination between episomal plasmids and chromosomes in yeast. Genetics 105:843–856

    Google Scholar 

  • Federoff HJ, Cohen JD, Eccleshall TR, Needleman RB, Buchferer BA, Giacalone J, Marmur J (1982) Isolation of a maltase structural gene from Saccharomyces carlsbergiensis. J Bacteriol 149:1064–1070

    Google Scholar 

  • Federoff HJ, Eccleshall TR, Marmur J (1983a) Regulation of maltase synthesis in Saccharomyces carlsbergiensis. J Bacteriol 156:1301–1308

    Google Scholar 

  • Federoff HJ, Eccleshall TR, Marmur J (1983b) Carbon Catabolite Repression of maltase synthesis in Saccharomyces carlsbergiensis. J Bacteriol 156:301–307

    Google Scholar 

  • Hollenberg CP (1982) Cloning with 2μm DNA vectors and the expression of foreign genes in Saccharomyces cerevisiae. In: Hofschneider PH, Goebel W (eds) Current topics in microbiology and immunology, vol 96. Springer Verlag Berlin Heidelberg New York, pp 119–144

    Google Scholar 

  • Khan NA, Zimmermann FK, Eaton NR (1973) Genetic control of maltase formation in yeast. II. Evidence for a gene regulating the level of maltase production. Mol Gen Genet 124:365–367

    Google Scholar 

  • McMaster GK, Carmichael GG (1977) Analysis of single- and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine organge. Proc Natl Acad Sci USA 74:4835–4838

    Google Scholar 

  • Michels CA, Needleman RB (1983) A genetic and physical analysis of the MAL1 and MAL3 standard strains of Saccharomyces cerevisiae. Mol Gen Genet 191:225–230

    Google Scholar 

  • Nasmyth KA, Reed SI (1980) Isolation of genes by complementation in yeast: Molecular cloning of a cell cycle gene. Proc Natl Acad Sci USA 77:2119–2123

    Google Scholar 

  • Naumov GI (1976) Comparative genetics of yeasts. XVI. Genes for maltose fermentation in Saccharomyces carlsbergiensis. Genetika 12:87–100

    Google Scholar 

  • Needleman RB, Michels CA (1983) A repeated family of genes controlling maltose fermentation in Sacharomyces carlsbergiensis. Mol Cell Biol 3:796–802

    Google Scholar 

  • Rigby PWJ, Dieckmann M, Rhodes C, Berg P (1977) Labeling deoxyribonucleic acid to high specific activity by nick translation with DNA polymerase I. J Mol Biol 113:237–251

    Google Scholar 

  • Schmitt HD, Ciriacy M, Zimmermann FK (1983) The synthesis of yeast pyruvate decarboxylase is regulated by large variations in the messenger RNA level. Mol Gen Genet 192:247–252

    Google Scholar 

  • Struhl K, Stinchcomb DT, Scherer S, Davis RW (1979) Highfrequency transformation of yeast: Autonomous replication of hybrid DNA molecules. Proc Natl Acad Sci USA 76:1035–1039

    Google Scholar 

  • Tautz D, Renz M (1983) An optimized freeze-squeeze method for the recovery of DNA fragments from agarose gels. Anal Biochem 132:14–19

    Google Scholar 

  • tenBerge AMA, Zoutewelle G, van de Poll KW (1973) Regulation of maltose fermentation in Saccharomyces carlsbergiensis. I. The function of the gene MAL6 as recognized by mal6-mutants. Mol Gen Genet 123:233–246

    Google Scholar 

  • Wahl GM, Stern M, Stark GR (1979) Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethylpaper and rapid hybridization by using dextran sulfate. Proc Natl Acad Sci USA 76:3683–3687

    Google Scholar 

  • Zamenhoff S (1958) In: Preparation and assay of desoxyribonucleic acids from animal tissue. Colowick SP, Kaplan NO (eds) Methods in enzymology, vol 3. Academic Press, London, New York, pp 702

    Google Scholar 

  • Zimmermann FK, Khan NA, Eaton NR (1973) Identification of new genes involved in disaccharide fermentation in yeast. Mol Gen Genet 123:29–41

    Google Scholar 

  • Zimmermann FK, Eaton NR (1974) Genetics of induction and catabolite repression of maltase synthesis in Saccharomyces cerevisiae. Mol Gene Genet 134:261–272

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by H. Böhme

Rights and permissions

Reprints and permissions

About this article

Cite this article

Rodicio, R., Schmitt, H.D., Heinisch, J. et al. A hybrid DNA sequence containing the replication origin of the multicopy yeast plasmid 2 μm circle and an additional repeated sequence can convert maltose-negative into maltose-positive strains. Mol Gen Genet 197, 491–496 (1984). https://doi.org/10.1007/BF00329948

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00329948

Keywords

Navigation