YAC Protocols

  • David Markie

Part of the Methods in Molecular Biology™ book series (MIMB, volume 54)

Table of contents

  1. Front Matter
    Pages i-xii
  2. Zoia Larin, Anthony P. Monaco, Hans Lehrach
    Pages 1-11
  3. John E. Collins, Sheila Hassock, Ian Dunham
    Pages 13-21
  4. Charlotte G. Cole, John E. Collins, Ian Dunham
    Pages 23-31
  5. Charlotte G. Cole, John E. Collins, Ian Dunham
    Pages 33-47
  6. Gary A. Silverman
    Pages 65-68
  7. Jiannis Ragoussis
    Pages 69-74
  8. Shawn P. Iadonato, Andreas Gnirke
    Pages 75-85
  9. Alison Coffey, Simon Gregory, Charlotte G. Cole
    Pages 97-114
  10. Sandro Banfi, Huda Y. Zoghbi
    Pages 115-121
  11. Anna Di Rienzo, Amy C. Peterson, Nelson B. Freimer
    Pages 123-129
  12. Donald J. Ogilvie, Louise A. James
    Pages 131-138
  13. Gillian Bates
    Pages 139-144
  14. Gary A. Silverman
    Pages 145-155
  15. Jiannis Ragoussis, Anthony P. Monaco
    Pages 157-166
  16. Jennifer W. McKee-Johnson, Roger H. Reeves
    Pages 167-186
  17. Karen Duff, Clare Huxley
    Pages 187-198

About this book

Introduction

Yeast artificial chromosomes (YACs) have their origins in the molecular genetic analysis of the yeast Saccharomyces cerevisiae. The construction of self-maintaining genetic elements from isolated frag­ ments of the yeast genome defined DNA sequences necessary for chro­ mosome function has provided telomeres, centromeres, and autonomous replicating sequences. In 1987 a reversal of the strategy put these short functional DNA sequences to work in cloning vectors, producing "yeast" chromosomes largely composed of foreign DNA. Initially the insert size of clones averaged several hundred kilobasepairs, a remarkable achieve­ ment. Rapid progress with cloning technology has since enabled the construction of YAC libraries with average insert sizes of around 1 Mb, with many clones exceeding that size, and YACs remain the largest capacity microbiological cloning system available. They effectively bridge the size gap between bacterial cloning (plasmids, cosmids, PI, and bacterial artificial chromosomes) and what could be considered mammalian cloning systems (somatic cell hybrids and irradiati- fusion gene transfer hybrids). YACs also brought with them a conceptual revolution in the man­ agement of clone libraries. The large carrying capacity of YACs, with subsequent reduction in the total number required, meant that it was conceivable to store clones individually rather than as pools that require constant re-plating. Each clone in the library has a unique address and, with successive screenings, information accumulates about individual clones.

Editors and affiliations

  • David Markie
    • 1
  1. 1.Paediatric Research UnitUnited Medical and Dental Schools of Guy’s and St. Thomas’ HospitalsLondon

Bibliographic information

  • DOI https://doi.org/10.1385/0896033139
  • Copyright Information Humana Press 1996
  • Publisher Name Humana Press
  • eBook Packages Springer Protocols
  • Print ISBN 978-0-89603-313-9
  • Online ISBN 978-1-59259-541-9
  • Series Print ISSN 1064-3745
  • Series Online ISSN 1940-6029
  • About this book