Bivalent DNA vaccine of type A and type O foot-and-mouth disease virus and its preparing process

A foot-and-mouth disease virus and DNA vaccine technology, which is applied in the field of DNA vaccines, can solve the problems of vaccines containing highly toxic pathogenic substances, large-scale epidemics of diseases, and short validity periods of vaccines, and achieves the advantages of being conducive to popularization and use, long validity periods, and no special requirements. Effect

Inactive Publication Date: 2004-12-29
XIAMEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although these two vaccines have good immunogenicity, they are produced with poor safety. It is necessary to take protective measures for the laboratory and the staff participating in the experiment to ensure that they are not contaminated by pathogenic substances; to ensure that the virus does not leak
During the vaccine production process, the virus may not be completely killed or sufficiently attenuated, which will cause the vaccine to contain highly toxic pathogenic substances, which will cause the immunized animals to become ill and cause a large-scale epidemic of the disease
Most current vaccines have a short validity period and often need to be frozen, which increases the cost of vaccine promotion and use in rural areas

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] The synthetic sequence comprising the coding sequence of the O-type foot-and-mouth disease virus VP1 protein antigenic determinant gene, the A-type foot-and-mouth disease virus VP1 protein antigenic determinant gene coding sequence, and the linking fragment is combined with the eukaryotic early promoter of the simian vacuolar virus SV40 after enzyme digestion The expression vectors were connected to transform Escherichia coli competent cells. Positive clones were identified by enzyme digestion and PCR, and then confirmed by sequencing. Place the positive clone in a resistant medium and culture overnight, add chloramphenicol the next day, continue shaking culture, collect the bacteria, suspend in the buffer, add freshly prepared NaOH and SDS to lyse the bacteria, add the cold water Neutralize the neutralizing solution, place it and centrifuge; filter the supernatant through gauze, add isopropanol, centrifuge, and wash the precipitate with ethanol; dissolve the precipitat...

Embodiment 2

[0038]Will comprise O-type foot-and-mouth disease virus VP1 protein antigenic determinant gene coding sequence, A-type foot-and-mouth disease virus VP1 protein antigenic determinant gene coding sequence, the synthetic sequence of linking fragment, after digesting with the true Rou's tumor virus promoter (RSV) The nuclear expression vector was connected to transform Escherichia coli competent cells. Positive clones were identified by enzyme digestion and PCR, and then confirmed by sequencing. Place the positive clone in a resistant medium and culture overnight, add chloramphenicol the next day, continue shaking culture, collect the bacteria, suspend in the buffer, add freshly prepared NaOH and SDS to lyse the bacteria, add the cold water Neutralize with the neutralizing solution, and centrifuge after standing. All the other steps are the same as in Example 1.

Embodiment 3

[0040] Will comprise O-type foot-and-mouth disease virus VP1 protein antigenic determinant gene coding sequence, type A foot-and-mouth disease virus VP1 protein antigenic determinant gene coding sequence, the synthetic sequence of linking fragment, digest with the eukaryotic main late promoter (ADV) of adenovirus The expression vectors were connected, and the competent cells of Escherichia coli were transformed the next day. Positive clones were identified by enzyme digestion and PCR, and then confirmed by sequencing. Place the positive clone in a resistant medium and culture overnight, add chloramphenicol the next day, continue shaking culture, collect the bacteria, suspend in the buffer, add freshly prepared NaOH and SDS to lyse the bacteria, add the cold water Neutralize with the neutralizing solution, and centrifuge after standing. All the other steps are the same as in Example 1.

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PUM

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Abstract

The present invention relates to one kind of bivalent livestock type-A and type-O foot and mouth disease DNA vaccine. It is prepared through connecting serially the coding regions of the VP1 protein antigen determinants of both type-A and type-O foot and mouth disease virus strains, and the connection with DNA segment capable of raising the immunogenicity. The preparation process includes connecting enzyme incised segment with eukaryotic expression carrier and transforming competent colibacillus cell; PCR identification to obtain positive cloning; culturing, collecting thallus, cracking cell and centrifuging; and other steps. The bivalent livestock type-A and type-O foot and mouth disease DNA vaccine is used to immunize livestock to generate antibody resisting both type-A and type-O foot and mouth disease viruses. It has no pathogenetic effect and can induce the livestock body to generate comprehensive immune response and express modified natural antigen.

Description

technical field [0001] The invention relates to a DNA vaccine, in particular to a double-valent DNA vaccine of livestock type A and O foot-and-mouth disease virus and a preparation method thereof. Background technique [0002] Foot-and-mouth disease is a fast-spreading and highly infectious febrile infectious disease that causes artiodactyl animals. The International Office of Epizootics lists it as the first class A infectious disease. The pathogen of the disease is foot-and-mouth disease virus (foot-and-mouth disease, FMDV), which belongs to the family picornaviridae foot-and-mouth disease virus. Over the years, many countries have invested a lot of manpower, material resources and financial resources in order to prevent and control foot-and-mouth disease, but the disease is still prevalent in a wide range. [0003] Since the disease was discovered in 1989, people have developed attenuated vaccines and inactivated vaccines for foot and mouth disease virus, and have been w...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/135A61K39/295A61K48/00A61P31/14
Inventor 陈亮邵寒娟苏勇波林涛
Owner XIAMEN UNIV
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