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PRRSV and PCV-2bivalent recombinant fowl pox virus disease live vector vaccines

A live carrier vaccine, PCV-2 technology, applied in the direction of drug combination, antibody medical components, botanical equipment and methods, etc., can solve the problems of no vaccine, immunogenicity and safety limitations, and achieve good genetic stability Effect

Inactive Publication Date: 2008-01-30
金宁一
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

At present, there is no practical and effective method for the control of these two viral diseases. The vaccines for preventing PRRS mainly include inactivated vaccines and attenuated vaccines, but they are limited due to problems such as immunogenicity and safety.
There is currently no vaccine for the control of PCV-2

Method used

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  • PRRSV and PCV-2bivalent recombinant fowl pox virus disease live vector vaccines
  • PRRSV and PCV-2bivalent recombinant fowl pox virus disease live vector vaccines
  • PRRSV and PCV-2bivalent recombinant fowl pox virus disease live vector vaccines

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Embodiment Construction

[0023] The invention amplifies the ORF5 and ORF3 genes of PRRSV Changchun strain and the ORF2 gene of PCV2 Inner Mongolia strain by PCR method, clones them into pMD18-T vector, and constructs pMD18T-ORF2, pMD18T-ORF5 and pMD18T-ORF3 plasmids. The PRRSV structural protein gene ORF5-ORF3 was inserted into the downstream of the PUTAL composite promoter ATI-P7.5×20 of the fowlpox virus expression vector, and the PCV capsid protein gene ORF2 was inserted into the downstream of the tandem promoter P7.5×16 to construct respectively The recombinant fowlpox virus expression plasmid containing ORF5, ORF3 and ORF2 of PRRS was obtained; the recombinant plasmid and the fowlpox virus strain were co-transfected into chicken embryo fibroblasts for homologous recombination, and 5-bromo-2 - Deoxyuridine (Brdu) was screened under pressure; the recombinant virus was detected by polymerase chain reaction (PCR), Western blot (Western blot) and immunofluorescence test respectively, and the positive r...

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Abstract

The present invention relates to a PRRSV and PCV-2 bivalent recombinant fowlpox live vector vaccine, which pertains to the field of biotechnology. The present invention aims at providing the PRRSV and PCV-2 bivalent recombinant fowlpox live vector vaccine which has ORF5, ORF3 genes to express the structural glycoprotein of the PRRSV and ORF2 gene to express PCV-2 nucleocapsid protein and can be used as the live vector vaccine for the prevention of PRRS and PCV-2 infection in our country. The present invention constructs the pMD18T-ORF2, pMD18T-ORF5 and pMD18T-ORF3 plasmids, the plasmids are inserted to the downstream of the compound promoter ATI-P7.5 multiplied by 20 of the fowlpox virus expression vector pUTAL, at the same time, the ORF2 gene of the PCV2 are inserted to the downstream of the linking promoter P7.5 multiplied by 16, and the present invention further constructs the recombinant fowlpox virus gene transfer plasmid pUTAL-ORF2-ORF5-ORF3 which contains the ORF3-ORF3 gene of the PRRSV and the ORF2 gene of the PCV2. The present invention can be used as the live vector vaccine for the prevention of PRRS-PCV2 in our country.

Description

Technical field: [0001] The invention belongs to the field of biotechnology. Background technique: [0002] PRRS (Porcine Reproductive and Respiratory Syndrome) is one of the severe infectious diseases of pigs caused by PRRSV (Porcine Reproductive and Respiratory Syndrome Virus). It is listed as a B-type animal infectious disease by the International Veterinary Bureau, mainly causing reproductive disorders in sows (such as abortion, premature birth, stillbirth, mummified fetus) and respiratory symptoms in piglets. [0003] There are currently two serotypes of PRRS occurring in the world: the European type and the American type. So far only the American type is popular in our country. Because PRRS mainly causes sow reproductive barriers, it has caused huge economic losses to my country's pig industry. [0004] PRRSV Arteriviridae and Arterivirus genus are non-segmented single-stranded positive-sense RNA viruses. The genome is approximately 15Kb long, is infectious, and co...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K39/275A61P11/00A61P15/00C12N15/863
Inventor 金宁一沈国顺李昌金扩世鲁会军金洪涛李霄田明尧
Owner 金宁一
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