Construction and characterization of a yeast artificial chromosome library containing seven haploid human genome equivalents.

DOI Web Site 被引用文献8件
  • H M Albertsen
    Centre d'Etude du Polymorphisme Humain, Paris, France.
  • H Abderrahim
    Centre d'Etude du Polymorphisme Humain, Paris, France.
  • H M Cann
    Centre d'Etude du Polymorphisme Humain, Paris, France.
  • J Dausset
    Centre d'Etude du Polymorphisme Humain, Paris, France.
  • D Le Paslier
    Centre d'Etude du Polymorphisme Humain, Paris, France.
  • D Cohen
    Centre d'Etude du Polymorphisme Humain, Paris, France.

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<jats:p>Prior to constructing a library of yeast artificial chromosomes (YACs) containing very large human DNA fragments, we performed a series of preliminary experiments aimed at developing a suitable protocol. We found an inverse relationship between YAC insert size and transformation efficiency. Evidence of occasional rearrangement within YAC inserts was found resulting in clonally stable internal deletions or clonally unstable size variations. A protocol was developed for preparative electrophoretic enrichment of high molecular mass human DNA fragments from partial restriction digests and ligation with the YAC vector in agarose. A YAC library has been constructed from large fragments of DNA from an Epstein-Barr virus-transformed human lymphoblastoid cell line. The library presently contains 50,000 clones, 95% of which are greater than 250 kilobase pairs in size. The mean YAC size of the library, calculated from 132 randomly isolated clones, is 430 kilobase pairs. The library thus contains the equivalent of approximately seven haploid human genomes.</jats:p>

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