Integrative selection of human chromosome-specific yeast artificial chromosomes.
Human specific "integrative selection vectors" (ISVs) were designed to optimize integration of a yeast-selectable marker specifically into yeast artificial chromosomes (YACs) derived from human but not mouse DNA. ISVs were transformed into a YAC genomic library constructed from DNA of a human-mouse somatic cell hybrid containing chromosome 21 (HSA21) as the only human chromosome. One percent of the yeast in the original library contained HSA21-derived YACs; between 45% and 54% of the yeast recovered after transformation with ISV vectors contained human YACs. Integrative selection provides a rapid means of obtaining a highly enriched population of human chromosome-specific YACs by eliminating the labor-intensive steps of isolating and screening primary transformants. The procedure is biased toward the selection of YACs that contain a large number of targets for homologous recombinations; thus, libraries constructed by this procedure will be composed primarily of the largest YACs in the population.
Pavan
W J
WJ
Department of Physiology, Johns Hopkins University School of Medicine, Baltimore, MD 21205.
Reeves
R H
RH
D015825:Chromosomes, Fungal; D002877:Chromosomes, Human; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D005822:Genetic Vectors; D006801:Humans; D010957:Plasmids; D012091:Repetitive Sequences, Nucleic Acid; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000235:genetics*
D004247:DNA