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Molecular and General Genetics MGG

, Volume 205, Issue 2, pp 353–357 | Cite as

Cloning of the ARO3 gene of Saccharomyces cerevisiae and its regulation

  • Sadao Teshiba
  • Rolf Furter
  • Peter Niederberger
  • Gerhard Braus
  • Gerhard Paravicini
  • Ralf Hütter
Article

Summary

Regulation of the two isozymes of 3-deoxy-D-arabino-heptulosonate-7-phosphate synthase (DAHP synthase; EC 4.1.2.15) encoded by the genes ARO3 and ARO4 of Saccharomyces cerevisiae was studied. Both genes were shown to respond equally well to the general control of amino acid biosynthesis. Strains with mutations in these two genes were obtained by selecting first for a single aro3 mutation and afterwards for a double aro3 aro4 mutation. Gene ARO3, coding for the phenylanine-dependent isozyme of DAHP synthase was cloned on the 2 μm multicopy vector pJDB207 by complementation of mutation aro3-1 in yeast. The ARO3 gene, carried originally on a 9.6 kb BamHI fragment (plasmid pME541A), was subcloned on a 1.9 kb HindIII-XbaI fragment (plasmid pME543). A transcript of about 1.5 kb was shown to proceed from the HindIII towards the XbaI site. Expression from the 9.6 kb as well as from the 1.9 kb fragment was normal on a multicopy vector, since in both cases DAHP synthase levels of about 50-fold the wild-type level were observed.

Key words

Saccharomyces cerevisiae ARO3 gene DAHP synthase Regulation Gene cloning 

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Copyright information

© Springer-Verlag 1986

Authors and Affiliations

  • Sadao Teshiba
    • 1
  • Rolf Furter
    • 1
  • Peter Niederberger
    • 1
  • Gerhard Braus
    • 1
  • Gerhard Paravicini
    • 1
  • Ralf Hütter
    • 1
  1. 1.Mikrobiologisches Institut, Eidgenössische Technische HochschuleETH-ZentrumZürichSwitzerland

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