Isolation of single-copy human genes from a library of yeast artificial chromosome clones

Bernard H. Brownstein, Gary A. Silverman, Randall D. Little, David T. Burke, Stanley J. Korsmeyer, David Schlessinger, Maynard V. Olson

Research output: Contribution to journalArticlepeer-review

345 Scopus citations

Abstract

A recently developed cloning system based on the propagation of large DNA molecules as linear, artificial chromosomes in the yeast Saccharomyces cerevisiae provides a potential method of cloning the entire human genome in segments of several hundred kilobase pairs. Most applications of this system will require the ability to recover specific sequences from libraries of yeast artificial chromosome clones and to propagate these sequences in yeast without alterations. Two single-copy genes have now been cloned from a library of yeast artificial chromosome clones that was prepared from total human DNA. Multiple, independent isolates were obtained of the genes encoding factor IX and plasminogen activator inhibitor type 2. The clones, which ranged in size from 60 to 650 kilobases, were stable on prolonged propagation in yeast and appear to contain faithful replicas of human DNA.

Original languageEnglish
Pages (from-to)1348-1351
Number of pages4
JournalScience
Volume244
Issue number4910
DOIs
StatePublished - 1989

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