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Rabbit viral haemorrhagic virus capsid protein gene recombination baculovirus and bacterin

A technology of rabbit viral hemorrhage and recombinant baculovirus, which is applied in the fields of viral peptides, antiviral agents, genetic engineering, etc., can solve the problem of unstable supply of non-immune rabbits, increased cost of tissue inactivated vaccines, and restrictions on rabbit viral hemorrhage In order to achieve good immune efficacy, good immunogenicity and good antigenicity,

Active Publication Date: 2008-07-09
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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Problems solved by technology

In recent years, with the rise of industries such as rabbit products and the increase in demand for rabbits such as other animal vaccines, the market demand for rabbit viral hemorrhagic disease vaccines has increased a lot, but non-immune vaccines used to prepare tissue inactivated vaccines The supply of rabbits is very unstable, which causes the cost of traditional tissue-inactivated vaccines to increase continuously. In addition, tissue-inactivated vaccines have various disadvantages such as the potential risk of spreading virulence. However, RHDV has not been successfully passed on cells so far, which seriously restricts The development and production of a safer and more stable rabbit viral hemorrhagic disease vaccine has turned people's research on rabbit viral hemorrhagic disease vaccine to the development of a genetically engineered vaccine that can not only play a protective role, but also solve biological safety problems.

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  • Rabbit viral haemorrhagic virus capsid protein gene recombination baculovirus and bacterin

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Embodiment Construction

[0039] 1. Amplification and T / A Cloning of Rabbit Viral Hemorrhagic Disease Virus Capsid Protein VP60 Gene

[0040] 1. Extraction of Rabbit Viral Hemorrhagic Disease Virus (RHDV) Total RNA

[0041] Add the water treated with DEPC (diethylpyrocarbonate) to the rabbit liver tissue with rabbit viral hemorrhage at a volume ratio of 1:10, grind it into a suspension, and put it into a 1.5mL EP tube treated with DEPC , repeated freezing and thawing 3 times at -20°C, centrifuged at 7200r / min for 20min on a high-speed refrigerated centrifuge, took 200μL of supernatant, added 800μL of Trizol (TaKaRa Company), oscillated and mixed, left at room temperature for 10min, added 210μL of chloroform After shaking vigorously, place at room temperature for 1 min, centrifuge at 10,000 r / min at 4°C for 15 min, take 750 μL of the upper aqueous phase, add 0.5 mL of isopropanol, mix well, place at -20°C for 15 min, centrifuge at 10,000 r / min at 4°C for 10 min, and remove Clear, slowly add 1 mL of 75%...

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Abstract

The invention relates to recombinant baculoviral of capsid protein of rabbit viral hemorrhagic disease viruses and gene engineering vaccines, which belong to the bio-pharmaceutical field. A capsid protein VP60 whole gene of the rabbit viral hemorrhagic disease viruses is amplified through RT-PCR, a baculoviral recombinant vector is constructed through the molecular biotechnology, Sf9 cells are transfected, which enables the VP60 gene to obtain expression, and then gene engineering vaccines of the rabbit viral hemorrhagic disease are prepared through taking the capsid protein VP60 of the rabbit viral hemorrhagic disease viruses of the recombinant adenovirus expression as antigen and adding corresponding adjuvant. The invention can provide the capsid protein gene engineering vaccines of the rabbit viral hemorrhagic disease with good immune effects and simple technology, which is used for preventing and controlling the rabbit viral hemorrhagic disease.

Description

[0001] Technical field The present invention relates to recombinant baculovirus and genetic engineering vaccine of capsid protein of rabbit viral hemorrhagic disease virus, belonging to the field of biopharmaceuticals. Background technique [0002] Rabbit viral hemorrhagic disease, also known as rabbit plague, is a rabbit infectious disease characterized by acute, highly contagious, and large-scale death caused by rabbit viral hemorrhagic disease virus (RHDV). Infected rabbits usually die within 48 to 72 hours , the main features are highly infectious, respiratory system hemorrhage, liver necrosis, visceral edema, congestion and hemorrhage and other changes, which have caused huge economic losses to the rabbit industry all over the world. The International Veterinary Bureau listed the disease as a Class B infectious disease, and the No. 96 Order of the Ministry of Agriculture of my country promulgated it as a Class II disease. The emergence of RHD is relatively sudden and the ...

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Application Information

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IPC IPC(8): C12N15/866C12N15/40C07K14/08A61K39/12A61P31/14
Inventor 王芳薛家兵范志宇胡波徐为中张则斌何孔旺
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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