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Nucleic Acids Research, 1991, Vol. 19, No. 5 997-1000
© 1991

GENOME STRUCTURE AND MAPPING

Detection of homologous recombination between yeast artificial chromosomes with overlapping inserts

Alessandra Cellini1, Rosa M. Lacatena1,2 and Glauco p. tocchini-Valentini3,*

1Istituto Guido Donegani, Unità di Monterotondo via E.Ramarini 32, 00015 Monterotondo, Rome, Italy 2University of Chicago, Department of Molecular Genetics and Cell Biology Chicago, IL 60637-1433, USA 3Istituto di Biologia Cellulare CNR, 00137 Rome, Italy

* To whom correspondence should be addressed

Received January 10, 1991. Accepted February 4, 1991.

We have developed a system which facilitates the detection of recombination between Yeast Artificial Chromosomes (YAC's) carrying homologous Inserts. The system consists of a classical YAC vector, a new YAC vector and two appropriately labelled yeast strains of opposite mating type. The new YAC vector diners in markers from the canonical YAC vector. To test whether homologous recombination takes place, phage lambda DNA was cloned in the two vectors to provide a region of homology. The two constructs were then introduced into yeast strains of opposite mating type in which the endogenous genes for the selective markers present in the vectors are not expressed. Artificial chromosomes obtained by meiotic recombination are detected In the spores resulting from the mating.


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