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Nucleic Acids Research, 1994, Vol. 22, No. 7 1208-1214
© 1994

MOLECULAR BIOLOGY

Genetic selection of meiotic and mitotic recombinant yeast artificial chromosomes

Françoise Rotomondo+ and Georges F. Carle*,+

Centre de Biochimie, Unité 273 INSERM, Université de Nice Sophia-Antipolis Pare Valrose, 06108 Nice Cedex 2, France

*To whom correspondence should be addressed at present address

Received December 22, 1993. Revised March 7, 1994. Accepted March 7, 1994.

We have developed a genetic screen for the isolation of larger or smaller recombinant yeast artificial chromosomes derived from overlapping YACs. Integration plasmlds were used to modify the TRP1 and URA3 auxotrophlc markers present respectively on the left and right vector arms of one of the parental YACs. Olplolds containing the two parental YACs were studied through melosis and mitosis. Tetrad analysis revealed the presence of melotlc recombinant YACs at a frequency comparable with what is expected for yeast DNA (about 3 kb/cM). More direct genetic selection of diploids on -TRP-LYS synthetic media in the presence of 5-fluoro-orotlc acid (5-FOA), led to the isolation of mltotlc recombinant YACs at a high frequency. Analysis of these yeast cells by pulsed-fleld gel electrophoresls, confirmed the loss of both parental artificial chromosomes, and the specific retention of a larger or smaller recombinant YAC.

+ Present address: URA 1462 du CNRS. Université de Nice Sophia-Antipolis, Faculté de Médecine, Chemin de Valombrose, 06107 Nice Cedex 2, France


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