Preparation method for avian pasteurella genome vaccine

A technology of Pasteurella avian and genome, which is applied in the field of preparation of bacterial vaccines, can solve problems such as the complex antigen structure of Pasteurella avian, large local and systemic reactions, and failure to obtain an ideal attenuated live vaccine.

Inactive Publication Date: 2012-04-04
HENAN UNIV OF SCI & TECH
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Problems solved by technology

[0004] At present, the vaccines used in my country to prevent poultry cholera caused by Pasteurella avian mainly include attenuated live vaccines and inactivated vaccines. Among them, attenuated live vaccines are the most studied type of attenuated live vaccines for all poultry infectious diseases in my country. In the 1980s, it reached the peak period of development, but due to the inherent problems of Pasteurella avium's complex antigen structure and easy mutation, an ideal attenuated live vaccine has not yet been obtained.
There are a series of problems in the existing commercially available attenuated live vaccines, such as low immune protection, short immune period, large local and systemic reactions, resulting in food loss and serious drop in egg production, and sometimes even partial vaccination. The death of poultry, as well as technical problems such as insufficient genetic stability of the strain, improper application may even cause outbreaks of fowl cholera, so the effect is not very satisfactory
The problems of low protection rate and short immunity period of inactivated vaccines are more serious than attenuated live vaccines, so it is imperative to develop new and effective avian pasteurellosis vaccines

Method used

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  • Preparation method for avian pasteurella genome vaccine
  • Preparation method for avian pasteurella genome vaccine

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Embodiment Construction

[0065] A kind of preparation method of avian Pasteurella genome vaccine, concrete steps are:

[0066] Step 1: Extraction of the Pasteurella avian genome

[0067] 1) Take 0.1 ml of Pasteurella avium, inoculate it into 10 ml of tryptone soybean broth medium, and incubate for 18 hours at 37°C with a rotation speed of 200 rpm to form a bacterial liquid for later use;

[0068] 2) Take 10 ml of the bacterial solution in step 1), put it into a 15 ml centrifuge tube, put it into a centrifuge at a speed of 8000 rpm, centrifuge for 2 minutes, and pour out the sediment in the 15 ml centrifuge tube For the clarified liquid in the upper layer, add 950 microliters of tris-ethylenediaminetetraacetic acid solution to a 15-ml centrifuge tube, mix well, and add 50 microliters of 10% dodecyl to a 15-ml centrifuge tube. Alkyl sodium sulfate solution, 0.1 g of proteinase K, and then put a 15 ml centrifuge tube in a water bath at 37°C for 1 hour and set aside;

[0069] 3) Add 150 microliters of 5...

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Abstract

The invention discloses a preparation method for an avian pasteurella genome vaccine. The preparation method comprises the following steps of: performing enzyme digestion on a target gene by using a restrictive incision enzyme Sau3AI; performing enzyme digestion on an eucaryon expression vector PcDNA 3.1(+) by using a restrictive incision enzyme BamHI; performing dephosphorylation on the eucaryonexpression vector PcDNA 3.1(+) which is subjected to enzyme digestion by using calf intestinal alkaline phosphatase; connecting an avian pasteurella genome with the eucaryon expression vector pcDNA3.1(+); transform into a colon bacillus competence JM83; extracting plasmids; and identifying through enzyme digestion to obtain the avian pasteurella genome vaccine. As proved by animal experiment detection, the avian pasteurella genome vaccine prepared with the method can be used for reducing the generation of avian pasteurellosis and reducing the invasion of avian pasteurella to birds; and the preparation method is simple and is easy to operate.

Description

technical field [0001] The invention relates to a preparation method of a bacterial vaccine, in particular to a preparation method of an avian Pasteurella genome vaccine. Background technique [0002] Pasteurella avian (also known as Pasteurella multocida) is the pathogenic bacterium of fowl cholera (also known as avian pasteurellosis), and the fowl cholera disease caused by this bacteria is one of the more common poultry infectious diseases in my country , almost all poultry can be infected with the disease, and the morbidity and mortality of poultry infected with the disease are very high, with severe diarrhea, acute onset, short course of disease, and fast death as the main features. The nationwide prevalence of the disease has seriously affected the healthy and stable development of my country's poultry industry and brought huge economic losses to poultry farmers. [0003] Genomic vaccines are recombinant eukaryotic expression plasmids with antigen genes, which are used ...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K39/102A61P31/04
Inventor 宫强程茗牛明福孙晓菲张爱国孙军杰曲宁
Owner HENAN UNIV OF SCI & TECH
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