Unit

UNIT 5.14 Transfer of YAC Clones to New Hosts by Karyogamy-Deficient Mating

  1. Forrest Spencer1,
  2. Giora Simchen2

Published Online: 1 MAY 2001

DOI: 10.1002/0471142905.hg0514s06

Current Protocols in Human Genetics

Current Protocols in Human Genetics

How to Cite

Spencer, F. and Simchen, G. 2001. Transfer of YAC Clones to New Hosts by Karyogamy-Deficient Mating. Current Protocols in Human Genetics. 6:5.14:5.14.1–5.14.11.

Author Information

  1. 1

    Johns Hopkins University School of Medicine, Baltimore, Maryland

  2. 2

    The Hebrew University, Jerusalem, Israel

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: AUG 1995

Abstract

This unit provides a protocol for moving yeast artificial chromosome (YAC) clones to new yeast hosts using basic microbial techniques and pulsed-field gel analysis. In contrast to other methods that can be used to transfer YAC clones, this requires neither optimization to achieve high-efficiency DNA-mediated transformation of chromosome-sized DNA nor specialized equipment for tetrad dissection and analysis. Instead, chromosome (YAC) transfer is selected in rare segregants (“YACductants”) from a yeast mating that is rendered incomplete in most cell pairings by the presence of a kar1 (karyogamy-deficient) mutation in either parental strain. The Basic Protocol in this unit details the transfer of a YAC clone from yeast strain AB1380 (host to nearly all existing YAC libraries) to YPH925, a strain with nonreverting genetic markers compatible with existing plasmid constructs useful in YAC modification.