Reporter assay screens for protein targets in
View Patent ↗The present invention comprises responsive promoters along with screening methods which make use of the promoters in order to identify anti-fungal substances.
1. An isolated hybrid polynucleotide comprising a S.cerevisiae YIL066W-A Promoter operably linked to a reporter; with the proviso that the reporter operably linked to the S.cerevisiae YIL066W-A Promoter, is not the YIL066W-A open reading frame.
2. An isolated polynucleotide of claim 1 wherein said promoter comprises the nucleotide sequence set forth in SEQ ID NO: 23; and/or the reporter comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs: 42-51.
3. An isolated vector comprising the hybrid polynucleotide of claim 1 .
4. An isolated host cell comprising the vector of claim 3 .
5. The hybrid polynucleotide of claim 1 wherein said reporter is a member selected from the group consisting of S.cerevisiae ADE2, S.cerevisiae LYS2, S.cerevisiae TRP1, S.cerevisiae LEU2, S.cerevisiae URA 3, S.cerevisiae HIS3, Aequorea victoria GFP mutant 3, Renilla luciferase, Photinus pyralis luciferase , and E.coli 1acZ.
6. The hybrid polynucleotide of claim 5 wherein said reporter comprises the nucleotide sequence set forth in a member selected from the group consisting of SEQ ID NOs: 42-51.
7. The hybrid polynucleotide of claim 1 wherein the reporter is a Pho5 conjugated Renilla luciferase.
8. The host cell of claim 4 wherein said hybrid polynucleotide is chromosomally integrated.
9. The host cell of claim 4 further comprising a baseline promoter operably linked to a reporter.