The relative merits of the tetO2 and tetO7 promoter systems for the functional analysis of heterologous genes in yeast and a compilation of essential yeast genes with tetO2 promoter substitutions.
We have generated a collection of yeast strains, each of which has an essential yeast gene under the control of the tetracycline-responsive, tetO, promoter. Screens using first-generation promoter-swap strains uncovered the non-specific responsiveness of the tetO7 promoter to a known human transcription factor (hIRF-1). Non-specific regulation was not observed with the tetO2 promoter. Reporter assays have been used to demonstrate this phenomenon. Subsequent efforts to generate a collection of tetracycline-regulatable strains have focused on the tetO2 promoter. These strains are available to the yeast community and can be used for functional genomics studies.
Wishart
Jill A
JA
Faculty of Life Sciences, Michael Smith Building, University of Manchester, Oxford Road, Manchester M13 9PT, UK.
Osborn
Michael
M
Gent
Manda E
ME
Yen
Kuangyu
K
Vujovic
Zorana
Z
Gitsham
Paul
P
Zhang
Nianshu
N
Ross Miller
J
J
Oliver
Stephen G
SG
D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics*; D004271:DNA, Fungal / Q000235:genetics; D005800:Genes, Fungal / Q000235:genetics* / Q000502:physiology; D006801:Humans; D050856:Interferon Regulatory Factor-1 / Q000235:genetics; D007763:Lac Operon; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D016415:Sequence Alignment
D001426:Bacterial Proteins; D002352:Carrier Proteins; D004271:DNA, Fungal; D050856:Interferon Regulatory Factor-1; C083139:Tet O resistance protein, Bacteria