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Science 16 June 1989:
Vol. 244. no. 4910, pp. 1348 - 1351
DOI: 10.1126/science.2544027
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Articles

Science, Vol 244, Issue 4910, 1348-1351
Copyright © 1989 by American Association for the Advancement of Science

articles

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

BH Brownstein, GA Silverman, RD Little, DT Burke, SJ Korsmeyer, D Schlessinger, and MV Olson

Department of Genetics, Washington University School of Medicine, St. Louis, MO 63110.

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 application 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.


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