Current Genetics

, Volume 3, Issue 3, pp 173–180 | Cite as

Construction of new yeast vectors and cloning of the nif (nitrogen fixation) gene cluster of Klebsiella pneumoniae in yeast

  • Claude Gerbaud
  • Claudine Elmerich
  • Nicole Tandeau de Marsac
  • Patrick Chocat
  • Nicole Charpin
  • Michel Guérineau
  • Jean-Paul Aubert
Article

Summary

Two vectors, termed pG63.11 (7.6 Kb) and pHCG3 (9.6 Kb), suitable for yeast transformation have been constructed. The pHCG3 vector has cosmid properties. Both vectors contain a single 3.3 Kb EcoRI-HindIII fragment of yeast origin which carries the yeast URA3 gene (1.1 Kb) and the origin of replication of the 2 µm plasmid (2.2 Kb). They confer ampicillin resistance and they contain 5 unique EcoRI,HpaI,HindIII,BamHI and SalI restriction sites. Cosmid pHCG3 was used to clone the nitrogen fixation (nif) gene cluster of Klebsiella pneumoniae carried by twoHindIII fragments of 17 and 26 Kb, respectively. The resulting cosmid, termed pGPC875 (53 Kb) which conferred a Nif+ phenotype to Escherichia coli, was introduced in yeast by transformation. No acetylene reduction activity was detectable in the transformants. However it was shown that the entire information for nitrogen fixation can be replicated and maintained intact in yeast for more than 50 generations of growth.

Key words

Saccharomyces cerevisiae Yeast vectors Cosmids nif genes 

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Copyright information

© Springer-Verlag 1981

Authors and Affiliations

  • Claude Gerbaud
    • 1
  • Claudine Elmerich
    • 2
  • Nicole Tandeau de Marsac
    • 2
  • Patrick Chocat
    • 2
  • Nicole Charpin
    • 2
  • Michel Guérineau
    • 1
  • Jean-Paul Aubert
    • 2
  1. 1.Laboratoire de Biologie Génétique et MoléculaireUniversité de Paris SudOrsayFrance
  2. 2.Unité de Physiologic Cellulaire, Département de Biochimie et Génétique MoléculaireInstitut PasteurParis Cedex 15France

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