Leafhopper ecdysone receptor nucleic acids, polypeptides, and uses thereof
View Patent ↗The present invetion relates to a novel isolated leafhopper ecdysone receptor polypeptide. The invention also realtes 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 eypression in an ecdysone receptor-based gene expression modulation system and methods for identifying molecules that modulate leafhopper ecdysone receptor activity.
1. An isolated polynucleotide comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1, nucleotides 448-1109 of SEQ ID NO: 1, nucleotides 16-1109 of SEQ ID NO: 1, nucleotides 214-1109 of SEQ ID NO: 1, and nucleotides 418-1109 of SEQ ID NO: 1.
2. An isolated polynucleotide encoding a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 2, amino acids 150-370 of SEQ ID NO: 2, amino acids 6-370 of SEQ ID NO: 2, amino acids 72-370 of SEQ ID NO: 2, and amino acids 140-370 of SEQ ID NO: 2.
3. A vector comprising the isolated polynucleotide according to claims 1 or 2 .
4. The vector according to claim 3 , wherein the isolated polynucleotide is operatively linked to an expression control sequence that permits expression of the isolated polynucleotide in an expression competent host cell.
5. The vector according to claim 4 , wherein the expression control sequence comprises a promoter that is functional in a mammalian cell.
6. The vector according to claim 4 , wherein the vector is selected from the group consisting of a retroviral vector, plasmid, and a viral vector.
7. The vector according to claim 6 , wherein the vector is a plasmid.
8. An isolated host cell transfected with the vector according to claim 3 .
9. The isolated host cell according to claim 8 , wherein the host cell is selected from the group consisting of a bacterial cell, a fungal cell, a yeast cell, a nematode cell, an insect cell, a fish cell, a plant cell, an avian cell, an animal cell, and a mammalian cell.
10. An isolated polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 2, amino acids 150-370 of SEQ ID NO: 2, amino acids 6-370 of SEQ ID NO: 2, amino acids 72-370 of SEQ ID NO: 2, and amino acids 140-370 of SEQ ID NO: 2.
11. A method for producing a leafhopper ecdysone receptor polypeptide comprising:
a) culturing the host cell of claim 8 in culture medium under conditions permitting expression of a leafhopper ecdysone receptor polypeptide; and
b) isolating the leafhopper ecdysone receptor polypeptide from the culture medium.
12. A composition comprising the isolated polynucleotide according to claim 1 or 2 and a carrier.
13. A composition comprising the vector according to claim 3 and a carrier.
14. A composition comprising the isolated polypeptide according to claim 10 and a carrier.
15. A method of screening for molecules that modulate leafhopper ecdysone receptor activity in a cell, the method comprising:
a) contacting a cell comprising the isolated polypeptide according to claim 10 with a candidate molecule; and
b) detecting leafhopper ecdysone receptor activity in the presence of the molecule.
16. The method according to claim 15 , wherein the molecule is an agonist of leafhopper ecdysone receptor.
17. The method according to claim 15 , wherein the molecule is an antagonist of leafhopper ecdysone receptor.