ISSN:
1432-0983
Keywords:
Co-transformation
;
Gene cloning
;
DNA recombination
;
Plasmid replication
;
Aspergillus nidulans
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
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.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF00351716
Permalink