Expression cassette and host cell for expressing a Vip3-interacting protein
This invention provides polypeptides that were identified as interacting with Vip3 toxin. This invention also provides a method of identifying agents that bind to Vip3 interacting polypeptides, which agents may act as insecticidal agent, cytotoxic agents and/or modulate the activity of Vip3 interacting polypeptides.
1. An expression cassette comprising a promoter operably linked to a heterologous polynucleotide sequence that encodes a Vip3A-interacting protein from a Spodoptera frugiperda insect that is susceptible to a Vip3A insecticidal protein, wherein the Vip3A-interacting protein is a heat shock cognate 70 (Hsc70) protein comprising the amino acid sequence of SEQ ID NO: 3.
2. The expression cassette of claim 1 , wherein the promoter is functional in an insect cell.
3. The expression cassette of claim 2 , wherein the insect cell is a Spodoptera frugiperda cell.
4. The expression cassette of claim 3 , wherein the Spodoptera frugiperda cell is an Sf9 cell.
5. The expression cassette of claim 2 , wherein the insect cell is a Drosophila melanogaster cell.
6. The expression cassette of claim 5 , wherein the Drosophila melanogaster cell is an S2 cell.
7. A cultured insect cell comprising the expression cassette of claim 1 .
8. The cultured insect cell of claim 7 , wherein the cultured insect cell is a Spodoptera frugiperda cell.
9. The cultured insect cell of claim 8 , wherein the Spodoptera frugiperda cell is an Sf9 cell.
10. The cultured insect cell of claim 7 , wherein the cultured insect cell is a Drosophila melanogaster cell.
11. The cultured insect cell of claim 10 , wherein the Drosophila melanogaster cell is an S2 cell.
12. A cultured insect cell line comprising the cultured insect cell of claim 7 .
13. The cultured insect cell line of claim 12 , wherein the cultured insect cell line is a Spodoptera frugiperda Sf9 cell line.
14. The cultured insect cell line of claim 12 , wherein the cultured insect cell line is a Drosophila melanogaster S2 cell line.
15. A method of producing a Vip3A-interacting protein comprising culturing an insect cell comprising the expression cassette of claim 1 to thereby produce the Vip3A-interacting protein, wherein the Vip3A-interacting protein is a Hsc70 protein.