UNIT 5.14 Transfer of YAC Clones to New Hosts by Karyogamy-Deficient Mating
Published Online: 1 MAY 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
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.
- Published Online: 1 MAY 2001
- Published Print: AUG 1995
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.