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Applied and Environmental Microbiology, December 1999, p. 5303-5306, Vol. 65, No. 12
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Transformation of Escherichia coli with DNA from Saccharomyces cerevisiae Cell Lysates

Ana Cristina Adam, Gracia González-Blasco, Marta Rubio-Texeira,dagger and Julio Polaina*

Instituto de Agroquímica y Tecnología de Alimentos, Consejo Superior de Investigaciones Científicas, Apartado de Correos 73, E-46100 Burjassot, Valencia, Spain

Received 18 March 1999/Accepted 16 September 1999

We developed a system to monitor the transfer of heterologous DNA from a genetically manipulated strain of Saccharomyces cerevisiae to Escherichia coli. This system is based on a yeast strain that carries multiple integrated copies of a pUC-derived plasmid. The bacterial sequences are maintained in the yeast genome by selectable markers for lactose utilization. Lysates of the yeast strain were used to transform E. coli. Transfer of DNA was measured by determining the number of ampicillin-resistant E. coli clones. Our results show that transmission of the Ampr gene to E. coli by genetic transformation, caused by DNA released from the yeast, occurs at a very low frequency (about 50 transformants per µg of DNA) under optimal conditions (a highly competent host strain and a highly efficient transformation procedure). These results suggest that under natural conditions, spontaneous transmission of chromosomal genes from genetically modified organisms is likely to be rare.

* Corresponding author. Mailing address: Instituto de Agroquímica y Tecnología de Alimentos, Consejo Superior de Investigaciones Científicas, Apartado de Correos 73, E-46100 Burjassot, Valencia, Spain. Phone: 34-963 90 00 22. Fax: 34-963 63 63 01. E-mail: jpolaina{at}iata.csic.es.

dagger Present address: Kimmel Cancer Institute, Jefferson Medical College, Philadelphia, PA 19107.

Applied and Environmental Microbiology, December 1999, p. 5303-5306, Vol. 65, No. 12
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.




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