# US 7,745,175 B2 · Polynucleotides encoding Clostridium perfringens alpha toxin proteins

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 7,745,175 B2
- Document ID: US7745175B2
- Status: Granted
- Kind: B2
- Application number: 11/925,300
- Filed: 2007-10-26
- Published: 2008-05-01
- Granted: 2010-06-29
- Earliest claimed: 2006-10-27
- Assignees: Dow AgroSciences LLC
- Inventors: William M. Ainley; Janna Armstrong; Krishna Madduri; Donald J. Merlo; Kelley A. Smith; Mark A. Thompson; Steven R. Webb; Liu Y. Shen
- Web page: https://patentgrep.com/patent/US7745175B2

## CPC

- A61K 39/08: Clostridium, e.g. Clostridium tetani
- A61P 1/00: Drugs for disorders of the alimentary tract or the digestive system
- A61P 43/00: Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
- C07K 14/33: from Clostridium (G)
- A61K 2039/552: {Veterinary vaccine}
- A61K 2039/55577: {Saponins; Quil A; QS21; ISCOMS}
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- 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)

## Abstract

This invention pertains in part to the development of a vaccine for poultry against necrotic enteritis (NE). The vaccine utilizes a protective antigen that is a mutated, full-length, non-toxic Clostridium perfringens (Cp) α-toxin protein (Mcpa). Utility of this vaccine was demonstrated by reduction of lesion severity in NE challenge trails, for example. Also disclosed herein are novel approaches for producing this vaccine in significant quantities. One exemplified approach involves producing NE vaccine (mutated alpha toxin) in bacterial expression systems, preferably utilizing the Pseudomonas fluorescens system, for commercial use in controlling NE in the poultry industry. The subject vaccines can be administered preferably to chickens in several different ways. A novel, Type VI alpha toxin from chicken isolates of Cp is also disclosed.

## Claims

### Claim 1

A polynucleotide that encodes a protein comprising SEQ ID NO:4.

### Claim 2 (depends on 1)

The polynucleotide of claim 1 comprising a sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:7, SEQ ID NO:9, and SEQ ID NO:11.

### Claim 3 (depends on 1)

A method of producing a protein encoded by the polynucleotide of claim 1, said method comprising constructing a plant expression vector comprising a nucleic acid sequence that encodes said protein, transforming a plant cell with said plant expression vector, culturing said transformed plant cell under conditions suitable for production of said protein, and enriching said protein from said transformed plant cell.

### Claim 4 (depends on 1)

A method of producing protein encoded by the polynucleotide of claim 1, said method comprising constructing a prokaryotic expression vector comprising a polynucleotide that encodes said protein, transforming a prokaryotic cell with said prokaryotic expression vector, culturing said transformed prokaryotic cell under conditions suitable for production of said protein, and enriching said protein from said transformed prokaryotic cell.

### Claim 5 (depends on 4)

The method of claim 4 wherein said prokaryotic cell is a Pseudomonas fluorescens cell.

### Claim 6

An isolated polynucleotide that encodes a protein comprising residues 29 to 398 of SEQ ID NO:2.

### Claim 7 (depends on 6)

The polynucleotide of claim 6 that encodes SEQ ID NO:2.