ISSN:
1432-0983
Keywords:
Trans-kingdom conjugation
;
DNA integration
;
Saccharomyces cerevisiae
;
Escherichia coli
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Summary IncQ-derived conjugative shuttle vectors, which carried the yeast gene URA3 and/or the yeast autonomously replicating sequence (ARS1), were constructed. Both the ars-plus plasmid pAY205 and the ars-less plasmid pAY201 were successfully transmitted from E. coli to S. cerevisiae by the action of mob and tra. In this trans-kingdom conjugation, plasmid pAY205 could replicate and be retained in transconjugants. Plasmid pAY201 caused the formation of “micro-colonies” of abortive transconjugants due to its transient expression and rapid disappearance. Nevertheless, one per about 103 colonies caused by transmitted pAY201 plasmids were uncurable by integration into the homologous region of a yeast chromosome. Analyses by restriction enzyme mapping and Southern hybridization indicate that this integration is primarily caused by a double crossover during conjugation and not by a single reciprocal recombination.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF00318467
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