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.
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Communicated by K. Esser
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Nishikawa, M., Suzuki, K. & Yoshida, K. DNA integration into recipient yeast chromosomes by trans-kingdom conjugation between Escherichia coli and Saccharomyces cerevisiae . Curr Genet 21, 101–108 (1992). https://doi.org/10.1007/BF00318467
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DOI: https://doi.org/10.1007/BF00318467