Open Access Highly Accessed Methodology article

A strategy for constructing aneuploid yeast strains by transient nondisjunction of a target chromosome

Kirk R Anders*, Julie R Kudrna, Kirstie E Keller, BreAnna Kinghorn, Elizabeth M Miller, Daniel Pauw, Anders T Peck, Christopher E Shellooe and Isaac JT Strong

Author Affiliations

Biology Department, Gonzaga University, 502 E Boone Avenue, Spokane, WA 99258, USA

For all author emails, please log on.

BMC Genetics 2009, 10:36  doi:10.1186/1471-2156-10-36

Published: 13 July 2009

Additional files

Additional file 1:

Characterization of conditional centromere. (A) Conditional centromere does not cause growth defects on glucose. The heterozygous parent of KAY614, containing PGAL1-CEN3 and ura3::HIS3 at the CEN4 locus, was sporulated and tetrads were dissected onto YPD rich medium. (B) Kinetics of galactose-induced chromosome loss. A diploid strain, heterozygous for PGAL1-CEN3 URA3 at CEN3, was grown in YPD to log phase, washed and incubated in YP-galactose, plated to YPD, then phenotyped. The non-repressing sugar raffinose was used in later experiments instead of glucose [40], which is expected to allow more rapid induction of GAL1 promoter activity. (C) Galactose-induced loss of chromosome IV yields unstable 2N-1 phenotype. A diploid strain, heterozygous for PGAL1-CEN3 URA3 at CEN4, was grown overnight in YPD or YP-galactose, then plated to YPD. Most of the small colonies were Ura- and unstable, rapidly reverting to normal growth but remaining Ura-. This is consistent with endoreduplication of the remaining chromosome IV, as observed by Alvaro et al. [29].

Format: PDF Size: 69KB Download file

This file can be viewed with: Adobe Acrobat Reader

Open Data

Additional file 2:

Galactose induction of candidate chromosome VI disomes. Galactose induces the appearance of many small Ura+His+ papillae in a strain carrying a modified chromosome VI. Strain KAY628, harbouring the LEU2-marked TUB1 plasmid pKA55, was grown to log phase in raffinose-containing medium, split, and one-half was exposed to galactose for 1.3 culture doublings. 107 cells were spread to plates selecting for Ura+His+Leu+ papillae. Plates were incubated 3 days and photographed. Papillae were picked, colony-purified, then cultured for DNA isolation and array CGH as described in the text.

Format: JPEG Size: 1.8MB Download file

Open Data

Additional file 3:

Oligonucleotides used in this study. This table contains nucleotide sequences and genome coordinates of the oligonucleotides used for PCR.

Format: PDF Size: 161KB Download file

This file can be viewed with: Adobe Acrobat Reader

Open Data

Additional file 4:

Strain construction summaries. This file contains flowcharts that summarize the construction of yeast strains in this study.

Format: PDF Size: 126KB Download file

This file can be viewed with: Adobe Acrobat Reader

Open Data