Vaccine against necrotic enteritis in poultry

A poultry, vaccine technology, applied in] this application relates to the preparation of poultry necrotic enteritis, in another aspect, Clostridium perfringens pilus polypeptide, in one aspect provides an isolated gas-producing Clostridium perfringens fimbriae, polypeptides for vaccines, vaccines for necrotizing enteritis, say, immunogenicity field as described herein, able to resolve the strains that work, necrotizing enteritis non-specific, unknown necrotizing enteritis Pathogenesis and other issues

Pending Publication Date: 2020-01-14
AGRI & AGRI FOOD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In addition, many of these proteins are not specific to the strains that cause necrotizing enteritis and are not known to contribute to necrotizing enteritis pathogenesis

Method used

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  • Vaccine against necrotic enteritis in poultry
  • Vaccine against necrotic enteritis in poultry
  • Vaccine against necrotic enteritis in poultry

Examples

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example

[0068] Other features of the invention will become apparent from the following non-limiting examples, which illustrate by way of example the principles of the invention.

example 1

[0069] Example 1: Production of Purified Recombinant Pili-Associated Polypeptides from Clostridium perfringens

[0070] The coding regions of the three pilus subunits (cnaA, fimA and fimB) were codon optimized and truncated to exclude the predicted N-terminal signal peptide and C-terminal cell wall sorting signal LPXTG transmembrane domains. The C-terminal domain is a hydrophobic region predicted to be removed during pili assembly. Truncated codon-optimized coding regions were synthesized (Integrated DNA Technologies, Coralville, USA) using In-Fusion TM Cloning, cloned into pET28a expression vector (MilliporeSigma, Ontario, Canada) according to the manufacturer's instructions (Takara Bio USA, Mountain View, California, USA) Sequence verification was performed in Etobicoke, Ontario, Canada, and then transformed into E. coli BL21 cells. Transformed colonies were grown in 1 L of LB broth supplemented with 50 μg / ml kanamycin and 1 mM IPTG at 37° C. for 18 hours with shaking. Th...

example 2

[0082] Example 2: Preparation of Clostridium perfringens strain CP1 pili subunit knockout mutants

[0083] It is generally as described above (Yu Q. (Yu, Q.), Lepp D (Lepp, D.), Mehdizadeh Gohari I. (Mehdizadeh Gohari, I.), Wu T. (Wu, T.), Zhou H.(Zhou,H.), Yin X.(Yin,X.), Yu H.(Yu,H.), Prescott J.F.(Prescott,J.F.), Nie S.P.(Nie, S.P.), Xie M.Y. (Xie, M.Y.), Gong J. (Gong, J.), 2017, The pathogenesis of Clostridium perfringens-induced necrotic enteritis in poultry requires the Agr-like quorum sensing system (TheAgr-like quorum sensing system is required for necrotic enteritis pathogenesis in poultry caused by Clostridium perfringens), Infection and Immunity (Infection and Immunity) 85(6):e00975-16), induced by ClosTron mutation (Heap J.T. (Heap, J.T.) et al., Molecular Biol (Methods Mol.Biol.)(2010),646:165-182), the three pili subunit genes (cnaA, fimA and fimB) in the virulent Clostridium perfringens strain CP1 were inserted into ) were each inactivated to generate CP1 kno...

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Abstract

An immunogenic polypeptide selected from an isolated Clostridium perfringens pilus polypeptide, a variant of the pilus polypeptide; a fragment of the pilus polypeptide; and a fragment of the variant,is useful for the preparation of a vaccine for the treatment or prevention of enteric necrosis in poultry. The isolated Clostridium perfringens pilus polypeptide includes an assembled pilus or the pilus subunits CnaA, FimA and/or FimB.

Description

Background technique [0001] The present application relates to polypeptides suitable for the preparation of vaccines against poultry necrotic enteritis. More specifically, the present application relates to a Clostridium perfringens pili polypeptide suitable for use in the preparation of a vaccine against necrotic enteritis associated with Clostridium perfringens infection in poultry. [0002] Necrotic enteritis is an intestinal disease of poultry such as broiler chickens that cost the poultry industry an estimated $US 6 billion in 2015. Necrotic enteritis is mainly caused by certain type A strains of Clostridium perfringens that produce the NetB pore-forming toxin, which overgrows and adheres to the intestinal mucosa, eventually producing the characteristic lesions of the disease. Clostridium perfringens is a normal resident of the intestinal tract and typically only those strains carrying the NetB toxin are capable of causing necrotic enteritis. Necrotic enteritis is prima...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/33A61K39/08A61P31/04A61P37/04C07K16/12C12N15/31C12N15/85G01N33/569
CPCA61K39/08C07K14/33A61P31/04A61P37/04G01N33/56911G01N2333/33C07K16/1282A61K2039/552A61K2039/55577A61K2039/70
Inventor 龚建华D·莱普
Owner AGRI & AGRI FOOD
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