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A novel yeast genomic DNA library on a geneticin‐resistance vector
Author(s) -
Jauert Peter A.,
Jensen Linnea E.,
Kirkpatrick David T.
Publication year - 2005
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.1250
Subject(s) - biology , selectable marker , genomic dna , genomic library , genetics , multiple cloning site , auxotrophy , complementation , shuttle vector , saccharomyces cerevisiae , dna , library , computational biology , vector (molecular biology) , phenotype , yeast , mutant , gene , plasmid , base sequence , 16s ribosomal rna , recombinant dna
Abstract We have constructed a Saccharomyces cerevisiae genomic library utilizing a novel centromere‐containing vector bearing the TRP1 and KANMX4 G418‐resistance selectable markers. Random Sau 3AI genomic DNA fragments with an average size of 6.4 kb were introduced into a unique Bam HI site in the vector's polylinker. Six independent library pools were generated and the characteristics of these pools were determined. This library can be used to introduce low‐copy genomic DNA fragments into any strain that is not G418‐resistant, making it useful for low‐copy overexpression analysis or complementation of mutant phenotypes in strains that lack a suitable auxotrophy marker or only exhibit phenotypes on rich, undefined, growth media. Copyright © 2005 John Wiley & Sons, Ltd.