# US 11,028,407 B2 · Insecticidal combinations of polypeptides having improved activity spectrum and uses thereof

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- Number: US 11,028,407 B2
- Document ID: US11028407B2
- Status: Granted
- Kind: B2
- Application number: 16/092,341
- Filed: 2017-04-19
- Published: 2020-05-07
- Granted: 2021-06-08
- Earliest claimed: 2016-04-19
- Assignees: Pioneer Hi-Bred International, Inc.; E. I. Du Pont de Nemours and Company
- Applicants: Pioneer Hi-Bred International, Inc.; E. I. Du Pont De Nemours and Company
- Inventors: Albert L Lu; Gusui Wu
- Web page: https://patentgrep.com/patent/US11028407B2

## CPC

- C12N 15/8286: {for insect resistance}
- C07K 14/325: Bacillus thuringiensis crystal peptides, i.e. delta-endotoxins
- Y02A 40/146: Genetically Modified [GMO] plants, e.g. transgenic plants
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE

## Abstract

The disclosure provides nucleic acids, and variants and fragments thereof, derived from strains of Bacillus thuringiensis encoding variant polypeptides having increased pesticidal activity against insect pests, including Lepidoptera and Coleopteran. Particular embodiments of the disclosure provide isolated nucleic acids encoding pesticidal proteins, pesticidal compositions, DNA constructs, and transformed microorganisms and plants comprising a nucleic acid of the embodiments. These compositions find use in methods for controlling pests, especially plant pests.

## Claims

### Claim 1

A DNA construct comprising a nucleic acid molecule encoding a variant Cry1B polypeptide having at least 95% sequence identity to SEQ ID NO: 29 and having insecticidal activity; and a nucleic acid molecule encoding a PtIP-83 polypeptide having at least 95% sequence identity to SEQ ID NO: 62, 64, or 66 and having insecticidal activity.

### Claim 2 (depends on 1)

The DNA construct of claim 1, wherein the nucleic acid molecules encoding the variant Cry1B polypeptide and the PtIP-83 polypeptide are each operably linked to a heterologous regulatory element.

### Claim 3

A transgenic plant comprising a molecular stack of a nucleic acid molecule encoding a variant Cry1B polypeptide having at least 95% sequence identity to SEQ ID NO: 29 and having insecticidal activity; and a nucleic acid molecule encoding a PtIP-83 polypeptide having at least 95% sequence identity to SEQ ID NO: 62, 64, or 66 and having insecticidal activity.

### Claim 4 (depends on 3)

The transgenic plant of claim 3, wherein the nucleic acid molecules encoding the variant Cry1B polypeptide and the PtIP-83 polypeptide are each operably linked to a heterologous regulatory element.

### Claim 5

A transgenic plant comprising a breeding stack of a nucleic acid molecule encoding a variant Cry1B polypeptide having at least 95% sequence identity to SEQ ID NO: 29 and having insecticidal activity; and a nucleic acid molecule encoding a PtIP-83 polypeptide having at least 95% sequence identity to SEQ ID NO: 62, 64, or 66 and having insecticidal activity.

### Claim 6 (depends on 5)

The transgenic plant of claim 5, wherein the nucleic acid molecule encoding the variant Cry1B polypeptide and the PtIP-83 polypeptide are each operably linked to a heterologous regulatory element.

### Claim 7 (depends on 1)

A transgenic plant or progeny thereof comprising the DNA construct of claim 1, wherein said transgenic plant is corn or soy.

### Claim 8 (depends on 3)

The transgenic plant or progeny thereof comprising the molecular stack of claim 3, wherein said transgenic plant is corn or soy.

### Claim 9 (depends on 5)

The transgenic plant or progeny thereof comprising the breeding stack of claim 5, wherein said transgenic plant is corn or soy.

### Claim 10

A composition comprising a nucleic acid molecule encoding a variant Cry1B polypeptide having at least 95% sequence identity to SEQ ID NO: 29 and having insecticidal activity and a nucleic acid molecule encoding a PtIP-83 polypeptide having at least 95% sequence identity to SEQ ID NO: 62, 64, or 66 and having insecticidal activity.