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Nucleic Acids Research, 1994, Vol. 22, No. 16 3406-3411
© 1994

GENOME STRUCTURE AND MAPPING

Characterization of four human YAC libraries for clone size, chimerism and X chromosome sequence representation

Ramaiah Nagaraja, Juha Kere, Sandra MacMillan, Mochubeloa W.J. Masisi, Douglas Johnson, Barbara J. Molini, George R. Halley, Kim Wein, Michelle Trusgnich, Brian Eble, Brian Railey, Bernard H. Brownstein and David Schlessinger*

Center for Genetics in Medicine, Washington University School of Medicine, St Louis, MO 63110, USA

*To whom correspondence should be addressed

Received April 22, 1994. Revised July 25, 1994. Accepted July 25, 1994.

Four collections of human X-specific YACs, derived from human cells containing supernumerary X chromosomes or from somatic cell hybrids containing only X human DNA were characterized. In each collection, 80 – 85% of YAC strains contained a single X YAC. Five thousand YACs from the various libraries were sized, and cocloning was assessed in subsets by the fraction of YAC insert-ends with non-X sequences. Cocloning was substantial, ranging up to 50% for different collections; and in agreement with previous indications, in all libraries the larger the YACs, the higher the level of cocloning. In libraries made from human — hamster hybrid cells, expected numbers of clones were recovered by STS-based screening; but unexpectedly, the two collections from cells with 4 or 5 X chromosomes yielded numbers of YACs corresponding to an apparent content of only about two X equivalents. Thus it is possible that the DNA of inactive X chromosomes is poorly cloned into YACs, speculatively perhaps because of its specialized chromatin structure.


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