Modified vip3 polypeptides

Patent metadata

Field	Value
Publication number	US-2017291925-A1
Application number	US-201515506320-A
Country	US
Kind code	A1
Filing date	Aug 27, 2015
Priority date	Aug 29, 2014
Publication date	Oct 12, 2017
Grant date	—

Abstract

The present invention is directed to vegetative insecticidal proteins (Vips) modified to comprise heterologous carbohydrate binding modules and methods of use thereof.

First claim

1 . A modified Vip3 polypeptide comprising a heterologous carbohydrate binding module (CBM).

2 . The modified Vip3 polypeptide of claim 1 , wherein the heterologous CBM is substituted for all or a portion of Domain III of a Vip3 polypeptide.

3 . The modified Vip3 polypeptide of claim 1 , wherein the modified Vip3 polypeptide comprises all or a portion of Domain I and/or Domain II of a Vip3 polypeptide.

4 . The modified Vip3 polypeptide of claim 1 , wherein the modified Vip3 polypeptide comprises all or a portion of Domain IV of a Vip3 polypeptide.

5 . The modified Vip3 polypeptide of claim 1 , wherein the modified Vip3 polypeptide lacks all or a portion of Domain IV of a Vip3 polypeptide.

6 . The modified Vip3 polypeptide of claim 1 , wherein the modified Vip3 polypeptide comprises, in the amino terminal to carboxy terminal direction, all or a portion of Domain I of the Vip3 polypeptide, all or a portion of Domain II of the Vip3 polypeptide, the heterologous CBM, and optionally all or a portion of Domain IV of the Vip3 polypeptide.

7 . The modified Vip3 polypeptide of claim 1 , wherein the Vip3 polypeptide comprises all or a portion of any one of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, or SEQ ID NO:6.

8 . The modified Vip3 polypeptide of claim 1 , wherein all or a portion of Domain III comprises:
(a) amino acids 542 to 667 of SEQ ID NO:1; (b) amino acids 542 to 667 of SEQ ID NO:2; (c) amino acids 542 to 667 of SEQ ID NO:3; (d) amino acids 550 to 675 of SEQ ID NO:4; (e) amino acids 552 to 677 of SEQ ID NO:6; or (f) a corresponding amino acid sequence of another Vip3 polypeptide.

9 . The modified Vip3 polypeptide of claim 1 , wherein the modified Vip3 polypeptide has at least 75% amino acid sequence similarity with an amino acid sequence selected from any one of SEQ ID NOs:1-23 or SEQ ID NOs:33-56.

10 . The modified Vip3 polypeptide of claim 1 , wherein the heterologous CBM comprises all or a functional portion of a CBM amino acid sequence from a β-1,4-mannanase, optionally a Type B CBM amino acid sequence from a β-1,4-mannanase.

11 . The modified Vip3 polypeptide of claim 1 , wherein the heterologous CBM comprises a CBM amino acid sequence that has at least 75% amino acid sequence similarity with the amino acid sequence of any one of SEQ ID NOs:24-32.

12 . The modified Vip3 polypeptide of claim 1 , wherein the heterologous CBM comprises a metal binding site.

13 . The modified Vip3 polypeptide of claim 1 , wherein the modified Vip3 polypeptide is pesticidal.

14 . The modified Vip3 polypeptide of claim 1 , wherein the modified Vip3 polypeptide is pesticidal against an insect.

15 . The modified Vip3 polypeptide of claim 1 , wherein the modified Vip3 polypeptide is pesticidal against a lepidopteran insect.

16 . The modified Vip3 polypeptide of claim 1 , wherein the modified Vip3 polypeptide is pesticidal against at least one of Ostrinia nubilalis (European corn borer), Plutella xylostella (diamondback moth), Spodoptera frugiperda (fall armyworm), Agrotis ipsilon (black cutworm), Agrotis orthogonia (pale western cutworm), Striacosta albicosta (western bean cutworm), Helicoverpa zea (corn earworm), Heliothis virescens (tobacco budworm), Spodoptera exigua (beet armyworm), Helicoverpa punctigera (native budworm), Helicoverpa armigera (cotton bollworm), Manduca sexta (tobacco hornworm), Trichoplusia ni (cabbage looper), Pectinophora gossypiella (pink bollworm), Diatraea grandiosella (southwestern corn borer), Diatraea saccharalis (sugarcane borer), Elasmopalpus lignosellus (lesser cornstalk borer), Psuedoplusia includens (soybean looper), Anticarsia gemmatalis (velvetbean caterpillar), Plathypena scabra (green cloverworm), Homoeosoma electellum (sunflower head moth), and Cochylis hospes (banded sunflower moth), or any combination thereof.

17 . A composition comprising the modified Vip3 polypeptide of claim 1 in an agriculturally acceptable carrier.

18 . A nucleic acid molecule comprising a nucleotide sequence encoding the polypeptide of claim 1 .

19 . The nucleic acid molecule of claim 18 , wherein the nucleotide sequence is selected from the group consisting of a nucleotide sequence of SEQ ID NOs:7-23, SEQ ID NOs:33-56, and any combination thereof.

20 . The nucleic acid molecule of claim 18 , wherein the nucleotide sequence has been codon optimized for expression in a plant.

21 . The nucleic acid molecule of claim 18 , further comprising a heterologous promoter sequence operatively linked to the nucleotide sequence encoding the polypeptide.

22 . The nucleic acid molecule of claim 21 , wherein said promoter is a plant-expressible promoter.

23 . An expression cassette and/or recombinant vector comprising the nucleic acid molecule of claim 18 .

24 . A transgenic host cell comprising the nucleic acid molecule of claim 18 .

25 . The transgenic host cell of claim 24 , wherein the transgenic host cell is a transgenic plant cell or a transgenic bacterial cell.

26 . The transgenic host cell of claim 25 , wherein the transgenic plant cell is selected from the group consisting of a sorghum cell, a wheat cell, a sunflower cell, a tomato cell, a cole crop cell, a cotton cell, a rice cell, a soybean cell, a sugar beet cell, a sugar cane cell, a tobacco cell, a barley cell, an oilseed rape cell and a maize cell.

27 . A transgenic plant comprising the transgenic plant cell of claim 25 .

28 . The transgenic plant of claim 27 , wherein the transgenic plant is selected from the group consisting of a sorghum plant, a wheat plant, a sunflower plant, a tomato plant, a cole crop plant, a cotton plant, a rice plant, a soybean plant, a sugar beet plant, a sugar cane plant, a tobacco plant, a barley plant, an oilseed rape plant and a maize plant.

29 . The transgenic plant of claim 27 , wherein the plant is a maize plant or a soybean plant comprising a nucleic acid molecule that comprises a modified Vip3 polypeptide, wherein said modified Vip3 polypeptide is pesticidal against an insect.

30 . The transgenic plant of claim 29 , wherein the modified Vip3 polypeptide is pesticidal against a lepidopteran insect.

31 . The transgenic plant of claim 29 , wherein the modified Vip3 polypeptide is pesticidal against at least one of Ostrinia nubilalis (European corn borer), Plutella xylostella (diamondback moth), Spodoptera frugiperda (fall armyworm), Agrotis ipsilon (black cutworm), Agrotis orthogonia (pale western cutworm), Striacosta albicosta (western bean cutworm), Helicoverpa zea (corn earworm), Heliothis virescens (tobacco budworm), Spodoptera exigua (beet armyworm), Helicoverpa punctigera (native budworm), Helicoverpa armigera (cotton bollworm), Manduca sexta (tobacco hornworm), Trichoplusia ni (cabbage looper), Pectinophora gossypiella (pink bollworm), Diatraea grandiosella (southwestern corn borer), Diatraea saccharalis (sugarcane borer), Elasmopalpus lignosellus (lesser cornstalk borer), Psuedoplusia includens (soybean looper), Anticarsia gemmatalis (velvetbean caterpillar), Plathypena scabra (green cloverworm), Homoeosoma electellum (sunflower head moth), and Cochylis hospes (banded sunflower moth), or any combination thereof.

32 . The transgenic plant of claim 29 , where

Assignees

• Watts Joseph M
• Syngenta Participations Ag

Inventors

• Watts Joseph M

Classifications

• C07K14/325

  Bacillus thuringiensis crystal peptides, i.e. delta-endotoxins · CPC title

• C12N15/8286

  for insect resistance · CPC title

• C07K2319/20

  containing a tag with affinity for a non-protein ligand · CPC title

• A01N37/46

  N-acyl derivatives · CPC title

• C07K14/195

  from bacteria · CPC title