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Disintegrator vectors for single‐copy yeast chromosomal integration
Author(s) -
Sadowski Ivan,
Su TingCheng,
Parent Jennifer
Publication year - 2007
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/yea.1469
Subject(s) - biology , plasmid , gene , genetics , computational biology , reporter gene , expression cassette , vector (molecular biology) , gene expression , recombinant dna
Vectors were developed for two‐step chromosomal integration of reporter genes or expression constructs. With these vectors, integration produces a disruption of the ADE8, LYS2, MET15, LEU2, HIS3 or FCY1 genes, and integrants can be easily identified by replica‐plating on selective media. Integration using these ‘disintegrator’ vectors produces a single‐copy integration of the construct of interest at the junction of the marker deletion, and removes the additional plasmid sequences. Importantly, the integrated constructs do not contain flanking sequence duplications, and therefore should be highly stable. Each of the vectors was shown to reliably integrate a TEF1–KAN expression cassette and/or GAL1–HIS3 and STE12–LacZ reporter genes. Copyright © 2007 John Wiley & Sons, Ltd.