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A kind of muscovy duck parvovirus VP3 gene recombinant fowlpox virus transfer vector and its construction method

A technology of muscovy duck parvovirus and transfer vector, which is applied in genetic engineering, plant genetic improvement, botanical equipment and methods, etc., can solve the problems of affecting the immune prevention effect of young muscovy ducks, the virulence of the attenuated vaccine is strong, and achieves the improvement Efficiency, improve expression efficiency, promote the effect of stable expression

Active Publication Date: 2019-03-26
INST OF ANIMAL HEALTH GUANGDONG ACADEMY OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The attenuated vaccine is easily interfered by the maternal antibody level, which affects the immune prevention effect of young muscovy ducks, and the attenuated vaccine also has the risk of returning to strong virulence

Method used

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  • A kind of muscovy duck parvovirus VP3 gene recombinant fowlpox virus transfer vector and its construction method
  • A kind of muscovy duck parvovirus VP3 gene recombinant fowlpox virus transfer vector and its construction method
  • A kind of muscovy duck parvovirus VP3 gene recombinant fowlpox virus transfer vector and its construction method

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Experimental program
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Effect test

Embodiment 1

[0052] 1 material

[0053] 1.1 Virus strains and cells

[0054] Muscovy duck parvovirus MDPV isolates were preserved in our laboratory, chickenpox virus FPV attenuated vaccine was purchased from Guangdong Yongshun Biopharmaceutical Co., Ltd.; chicken embryo fibroblasts were purchased from 9-day-old SPF chickens of Guangdong Dahuanong Animal Health Products Co., Ltd. Embryo preparation.

[0055] Strains and plasmids

[0056] The host strain DH5α was purchased from Beijing Tiangen Bioengineering Technology Service Co., Ltd., pMD18-T was purchased from TAKARA Company, the plasmids pBluescript II SK(+) and pcDNA3.1-EGFP were preserved in our laboratory, and the plasmid pMD18-VP3 was constructed by our laboratory . The early and late promoter P7.5 and late promoter P11 sequences of vaccinia virus were kindly donated by Professor Chen Zhiwei of the University of Hong Kong.

[0057] main reagent

[0058] Ex Taq DNA polymerase was purchased from TAKARA Company; T4 DNA ligase, res...

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Abstract

The invention discloses a Muscovy duck parvovirus VP3 genetic recombination fowl pox virus transfer vector and a building method thereof. The method includes: fowl pox virus genome is used as the template to amplify the homologous recombination arms TKL and TKR of TK genes, the TKL and TKR are connected into a TK fragment with multiple cloning sites (MCS), and the TK fragment is connected into a framework plasmid to obtain the plasmid pTK; a reverse serial connection expression box S is synthesized and inserted into the plasmid pTK to obtain the plasmid pTKS; VP3 genes are inserted into an MSC1 position behind an early promotor P7.5, and EGFP BGH pA is inserted into an MCS2 position behind a late promotor P11 to obtain the target transfer vector pTKS-VP3-EGFP. After the pTKS-VP3-EGFP transfects chicken embryonic fibroblast infected by the fowl pox virus, RT-PCR and fluorescent protein detection show that VP3 can be expressed normally. By the Muscovy duck parvovirus VP3 genetic recombination fowl pox virus transfer vector and the building method thereof, a foundation is laid on the further development of safe and efficient MDPV recombinant fowl pox virus vaccines.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a Muscovy duck parvovirus VP3 gene recombinant fowlpox virus transfer vector and a construction method thereof. Background technique [0002] Muscovy duck parvovirus disease is an acute, septic infectious disease caused by Muscovy duck parvovirus (MDPV), which is characterized by high infectivity and mortality. One of the serious infectious diseases. MDPV mainly affects ducklings less than 3 weeks old, also known as "three-week disease". The morbidity rate of young Muscovy ducks is 20% to 60%, and the fatality rate is 20% to 40%. The morbidity and mortality of Muscovy ducks over 30 days old are relatively low, and sick ducks often become stiff ducks. The occurrence of this disease is non-seasonal, and can occur throughout the year, and the emphasis is on prevention. At present, there is an attenuated vaccine to prevent the disease, which has good immunogenicity and genetic stability...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/863
Inventor 李林林胡奇林孙敏华董嘉文袁建丰邝瑞欢张建峰
Owner INST OF ANIMAL HEALTH GUANGDONG ACADEMY OF AGRI SCI
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