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Current Genetics

, Volume 24, Issue 6, pp 520–524 | Cite as

Co-transformation with autonomously-replicating helper plasmids facilitates gene cloning from an Aspergillus nidulans gene library

  • D. H. Gems
  • A. J. Clutterbuck
Original Articles

Abstract

Autonomously-replicating, marker-less “helper” plasmids were added to transformations of Aspergillus nidulans with plasmids which normally transform by chromosomal integration. This resulted in as much as a 200-fold increase in transformation efficiency. Recovery of autonomously-replicating plasmid co-integrates indicated that co-transformation involves recombination between integrating and helper plasmids, which occurs at a high frequency. Increasing DNA sequence-homology between pairs of plasmids used in simultaneous transformations enhanced co-transformation efficiency. Using helper plasmids and an A. nidulans gene library in a normally-integrating vector, the genes adC and adD were cloned as part of such a co-integrate. In effect, the addition of helper plasmid converts an integrating into an autonomously-replicating gene library in vivo.

Key words

Co-transformation Gene cloning DNA recombination Plasmid replication Aspergillus nidulans 

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Copyright information

© Springer-Verlag 1993

Authors and Affiliations

  • D. H. Gems
    • 1
  • A. J. Clutterbuck
    • 1
  1. 1.Institute of GeneticsGlasgow UniversityGlasgowScotland, UK

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