Clostridium perfringen alpha toxin genetic engineering vaccine and application thereof

Abstract

The invention relates to a clostridium perfringen alpha toxin genetic engineering vaccine and application thereof. The clostridium perfringen alpha toxin genetic engineering vaccine is prepared by designing a specific primer, implementing polymerase chain reaction (PCR) amplification on A-type clostridium perfringen DNA to obtain an alpha toxin gene segment, cloning the gene segment onto a pMD18-T Vector, selecting a positive recon and linking the positive recon onto a PET-28 alpha (+) plasmid; introducing the recombinant plasmid to a receptor strain to successfully construct a recombinant strain BL21 (DE3)-alpha; emulsifying the prepared alpha toxin protein and an aluminum hydroxide adjuvant at a volume ratio of 9:1, and adding 0.01% thiomersal to obtain the clostridium perfringen alpha toxin genetic engineering vaccine. The clostridium perfringen alpha toxin genetic engineering vaccine is easy for realization of industrial production, simple to operate and good in safety, and is capable of effectively preventing such diseases as animal necrotic enteritis, enterotoxemia, and human and animal traumatic gas gangrene caused by the A-type clostridium perfringens.

Description

(1) Technical field [0001] The invention discloses a Clostridium perfringens alpha toxin genetic engineering vaccine and its application, belonging to the technical field of animal genetic engineering. (2) Background of the invention [0002] Clostridium perfringens is one of the main pathogenic bacteria that cause various animal necrotic enteritis, enterotoxemia, human and animal traumatic gas gangrene and human food poisoning. The pathogenic factor of this bacteria is the exotoxin it produces. So far, there are 12 exotoxins (α, β, γ, δ, ε, η, θ, ι, κ, λ, μ, and ν) of Clostridium perfringens that have been found, but they play the main pathogenic role. There are only 4 species (α, β, ε and ι), according to which the bacteria are divided into 5 toxin types such as A, B, C, D and E. All types of Clostridium perfringens produce α-toxin, which is the main virulence factor of Clostridium perfringens. It has two enzyme activities, phospholipase C and sphingomyelinase, and can si...

AI Technical Summary

Problems solved by technology

These vaccines have a certain control effect on the epidemic of the disease, but they have the following disadvantages: the duration of the induced immune response is short, and multiple vaccinations are required; Different; due to the low level of induced immune response and the unbalanced vaccine response among the various antigenic components, it may induce disease; generally require the vaccine to be concentrated and purified; it cannot be vaccinated through natural routes, and it is not easy to produce local immune responses, etc., quality needs further improvement

[0004] The toxoid vaccine of alpha toxin has a protective effect in animals, but due to safety considerations, this vaccine is unlikely to be applicable to humans, so vaccine research must be based on a complete understanding of the structure and molecular function of the toxin, paying attention to the relationship between the toxin and the respiratory tract and gastrointestinal tract The interaction relationship of the track surface

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