Bacillus thuringiensis toxin receptors and uses thereof

Patent metadata

Field	Value
Publication number	US-9970926-B1
Application number	US-201414548905-A
Country	US
Kind code	B1
Filing date	Nov 20, 2014
Priority date	Nov 22, 2013
Publication date	May 15, 2018
Grant date	May 15, 2018

Abstract

The invention relates to identification and characterization of recombinant DNA and polypeptides for specific Bt toxin receptors. In particular, the Bt toxin receptors of the invention include those derived from the Lepidopteran super family including the species Trichoplusiani ni, Pseudoplusia includens, Helicoverpa zea , and Spodoptera frugiperda . The receptors of the invention further include those derived from the Coleopteran super family and particularly from the species Diabrotica virgifera virgifera . The recombinant DNA and polypeptides so provided are useful in the identification and design of novel Bt toxin receptor ligands including novel or improved insecticidal toxins for use in a variety of agricultural applications. Materials and methods for identifying novel toxins are also disclosed herein. The invention also provides methods for selecting toxins to combine to control insect populations by manipulating Bt toxin receptor.

First claim

What is claimed is:

1. A recombinant DNA molecule encoding a polypeptide derived from Spodoptera frugiperda having an amino acid sequence selected from the group consisting of:
a) an amino acid sequence as set forth in SEQ ID NO: 181;
b) an amino acid sequence having at least 90% sequence identity to the full length of the amino acid sequence set forth in SEQ ID NO: 181; and
c) an amino acid sequence having at least 95% sequence identity to the full length of the amino acid sequence set forth in SEQ ID NO: 181;
wherein said recombinant DNA molecule is operably linked to a heterologous promoter capable of initiating the transcription of the DNA sequence, and wherein said polypeptide has Bt toxin binding activity.

2. The recombinant DNA of claim 1 , wherein said recombinant DNA molecule has a nucleotide sequence of SEQ ID NO: 162.

3. A recombinant DNA vector comprising the recombinant DNA molecule according to claim 1 .

4. A non-human transgenic cell transformed with the recombinant DNA molecule according to claim 1 .

5. The transgenic cell of claim 4 , wherein said transgenic cell is an insect cell.

6. A non-human transgenic organism transformed with the recombinant DNA molecule according to claim 1 .

7. The transgenic organism according to claim 6 , wherein said transgenic organism is a prokaryotic or a eukaryotic organism.

8. The transgenic organism of claim 6 , wherein said transgenic organism is a whole insect.

9. A method to assess the toxicity of a candidate ligand, wherein said method comprises the steps of:
a) contacting said candidate ligand with cells comprising the recombinant DNA molecule of claim 1 and expressing said polypeptide with Bt toxin binding activity; and,
b) measuring the toxicity effect of said candidate ligand on said cells in terms of cell death indices.

Assignees

• Monsanto Technology Llc

Inventors

• Kraft Edward
• Bolognesi Renata
• Evdokimov Artem
• Moshiri Farhad
• Zheng Meiying
• Guzov Victor M

Classifications

• C07K16/28

  against receptors, cell surface antigens or cell surface determinants · CPC title

• G01N2500/04

  Screening involving studying the effect of compounds C directly on molecule A (e.g. C are potential ligands for a receptor A, or potential substrates for an enzyme A) · CPC title

• G01N33/5085 Primary

  of invertebrates · CPC title

• C07K16/40

  against enzymes · CPC title

• C12N9/16

  acting on ester bonds (3.1) · CPC title