RT Journal
A1 Kumar, Anuj
A1 Seringhaus, Michael
A1 Biery, Matthew C.
A1 Sarnovsky, Robert J.
A1 Umansky, Lara
A1 Piccirillo, Stacy
A1 Heidtman, Matthew
A1 Cheung, Kei-Hoi
A1 Dobry, Craig J.
A1 Gerstein, Mark B.
A1 Craig, Nancy L.
A1 Snyder, Michael
T1 Large-Scale Mutagenesis of the Yeast Genome Using a Tn7-Derived Multipurpose Transposon
JF Genome Research 
JO Genome Research 
YR 2004 
FD October 01 
VO 14 
IS 10a 
SP 1975 
OP 1986 
DO 10.1101/gr.2875304 
UL http://genome.cshlp.org/content/14/10a/1975.abstract 
AB We present here an unbiased and extremely versatile insertional library of yeast genomic DNA generated by in vitro mutagenesis with a multipurpose element derived from the bacterial transposon Tn7. This mini-Tn7 element has been engineered such that a single insertion can be used to generate a lacZ fusion, gene disruption, and epitope-tagged gene product. Using this transposon, we generated a plasmid-based library of ∼300,000 mutant alleles; by high-throughput screening in yeast, we identified and sequenced 9032 insertions affecting 2613 genes (45% of the genome). From analysis of 7176 insertions, we found little bias in Tn7 target-site selection in vitro. In contrast, we also sequenced 10,174 Tn3 insertions and found a markedly stronger preference for an AT-rich 5-base pair target sequence. We further screened 1327 insertion alleles in yeast for hypersensitivity to the chemotherapeutic cisplatin. Fifty-one genes were identified, including four functionally uncharacterized genes and 25 genes involved in DNA repair, replication, transcription, and chromatin structure. In total, the collection reported here constitutes the largest plasmid-based set of sequenced yeast mutant alleles to date and, as such, should be singularly useful for gene and genome-wide functional analysis.