Construction of a human DNA library in a circular centromere-based yeast plasmid.
The construction of a human DNA library in a centromere-based circular yeast plasmid is described. The vector contains the yeast CEN3 sequence, the URA3 gene for propagation in yeast and a hygromycin-resistance gene (HyR) for selection in mammalian cells. The library consists of 64,000 members with an average insert size of 150 kb, with some members containing inserts of > 1 Mb. We calculate that the library contains three human genome equivalents of DNA. Clones can be identified by a PCR-based screening of DNA pools from individual colonies that have been stored in microtiter wells.
McGonigal
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Pharmaceutical Products Division, Abbott Laboratories, Abbott Park, IL 60064, USA.
Bodelle
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Schopp
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Sarthy
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D001483:Base Sequence; D002503:Centromere / Q000235:genetics; D018244:Chromosomes, Artificial, Yeast / Q000235:genetics; D015723:Gene Library; D005822:Genetic Vectors; D006801:Humans; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics*