Leafthopper ecdysone receptor nucleic acids, polypeptides, and uses thereof
The present invention relates to a novel isolated leafhopper ecdysone receptor polypeptide. The invention also relates to an isolated nucleic acid encoding the leafhopper ecdysone receptor polypeptide, to vectors comprising them and to their uses, in particular in methods for modulating gene expression in an ecdysone receptor-based gene expression modulation system and methods for identifying molecules that modulate leafhopper ecdysone receptor activity.
1. A fusion protein comprising:
(a) a DNA-binding domain that is not an ecdysone receptor DNA-binding domain; and
(b) a homopteran ecdysone receptor ligand binding domain that is at least about 90% identical to amino acids 150-370 of SEQ ID NO: 2.
2. The fusion protein of claim 1 , wherein the homopteran ecdysone receptor is a leafhopper ecdysone receptor.
3. The fusion protein of claim 2 , wherein the leafhopper ecdysone receptor comprises the amino acid sequence of SEQ ID NO:2.
4. The fusion protein of claim 1 , wherein the DNA binding domain is a Gal4 DNA binding domain or a LexA DNA binding domain.
5. The fusion protein of claim 1 , further comprising a transactivation domain.
6. The fusion protein of claim 5 , wherein the transactivation domain is from a homopteran ecdysone receptor.
7. The fusion protein of claim 1 , wherein said homopteran ecdysone receptor ligand binding domain is at least about 95% identical to amino acids 150-370 of SEQ ID NO: 2.
8. The fusion protein of claim 1 , wherein said homopteran ecdysone receptor ligand domain is encoded by a polynucleotide sequence that is at least about 90% identical to nucleotides 448-1109 of SEQ ID NO: 1.
9. The fusion protein of claim 1 , wherein said homopteran ecdysone receptor ligand domain is encoded by a polynucleotide sequence that is at least about 95% identical to nucleotides 448-1109 of SEQ ID NO: 1.
10. The fusion protein of claim 1 , wherein said homopteran ecdysone receptor ligand domain is encoded by a polynucleotide sequence that hybridizes to nucleotides 448-1109 of SEQ ID NO: 1 in (a) a hybridization step of less than 500 mM salt and at least 37° Celsius, and (b) a washing step of 2×SSPE and at least 63° Celsius.
11. The fusion protein of claim 10 , wherein said homopteran ecdysone receptor ligand domain is encoded by a polynucleotide sequence that hybridizes to nucleotides 448-1109 of SEQ ID NO: 1 in (a) a hybridization step of less than 200 mM salt and at least 37° Celsius, and (b) a washing step of 2×SSPE and at least 63° Celsius.
12. The fusion protein of claim 1 , wherein said homopteran ecdysone receptor ligand domain is encoded by a polynucleotide sequence that hybridizes to nucleotides 448-1109 of SEQ ID NO: 1 in (a) a hybridization step of 2×SSPE at 63° Celsius, and (b) a washing step of 2×SSPE at 63° Celsius.
13. The fusion protein of claim 1 , wherein said homopteran ecdysone receptor ligand domain is encoded by a polynucleotide sequence that hybridizes to nucleotides 448-1109 of SEQ ID NO: 1 in (a) a hybridization step of 0.1×SSC and 0.1% SDS at 65° Celsius, (b) a washing step of 2×SSC and 0.1% SDS at 65° Celsius, and (c) a washing step of 0.1×SSC and 0.1% SDS at 65° Celsius.