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Recombinant pseudorabies virus, construction method and application thereof

A technique for recombining pseudorabies and viruses, which is applied in the direction of antiviral agents, virus antigen components, viruses/bacteriophages, etc., can solve the problems of raising pig raising costs, high production costs, and expensive prices, and achieve strong protection and improve expression efficiency , the effect of high proliferation titer

Inactive Publication Date: 2014-11-05
GUANGDONG WENS DAHUANONG BIOTECH +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the high production cost and high price of this kind of vaccine, the vaccine for piglets is 10 to 14 yuan per head. Based on the calculation of two immunizations, the cost of each piglet is at least 30 yuan, and the vaccine for sows is at least 30 yuan per head. The cost of immunization for 3 times is more than 90 yuan per pig. It can be seen that the use of this type of vaccine greatly increases the cost of raising pigs

Method used

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  • Recombinant pseudorabies virus, construction method and application thereof
  • Recombinant pseudorabies virus, construction method and application thereof
  • Recombinant pseudorabies virus, construction method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] The construction of rPRV-ORF2 (see the construction roadmap figure 1 )

[0049] 1) Treatment of diseased material and detection of PCV2: take 0.5-1g of lymph node tissue from 9 points of diseased pigs, cut them into granules with scissors, grind them in a grinder, add 1.5ml of normal saline, grind until homogenized, then transfer to 1.5ml In a sterilized centrifuge tube, 8000r / min, centrifuge for 2min, take the supernatant, extract the virus DNA according to the virus DNA extraction kit, and then use the primers PCV2F and PCV2R in Table 1 to detect. The PCR reaction system and reaction conditions are as follows:

[0050]

[0051] The results showed that 5 of the samples amplified the 493bp target band, which was PCV2-positive disease material;

[0052] 2) Amplification of the whole ORF2 gene and construction of pgE-ORF2 (see figure 2 ): Use the primers ORF2F and ORF2R in Table 1 to amplify the entire ORF2 gene from one of the PCV2-positive disease materials. The r...

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Abstract

The invention discloses a recombinant pseudorabies virus (PRV), a construction method and application thereof. The recombinant pseudorabies virus takes recombinant PRV / TK<-> / gE<-> / EGFP<+> as the female parent, an expression cassette containing the main immunogenic gene ORF2 of porcine circovirus type 2 is inserted to replace an EGFP gene expression cassette in a gE gene so as to obtain the recombinant pseudorabies virus named as rPRV-ORF2. The recombinant pseudorabies virus provided by the invention is used for recombinant pseudorabies virus vaccines and has safety and effectiveness.

Description

technical field [0001] The invention belongs to the technical field of animal virus genetic engineering, and in particular relates to a recombinant pseudorabies virus and its construction method and application. Background technique [0002] Pseudorabies virus (PRV) belongs to the Herpesvirus (Herpesviridae) alpha herpesvirus subfamily (Alpha-herpesvirinae) varicella virus (Varicello virus) type I (Porcine herpesvirus Type I), can cause severe disease in pigs . Pseudorabies is often an explosive epidemic in pigs, and it is particularly harmful to pregnant sows, often causing stillbirth, abortion, and mummies; gilts and empty sows infected with the virus can cause infertility, anoestrus, return to oestrus, and relapse. With infertility. After boars are infected, the testicles are swollen and atrophied, and they lose their seeding ability. A large number of newborn piglets die after being infected with PRV, the mortality rate can reach 100% within 15 days of age, and the mo...

Claims

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Application Information

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IPC IPC(8): C12N7/01C12N15/63A61K39/295A61K39/12A61K39/245A61P31/20A61P31/22C12R1/93
Inventor 黄红亮陈瑞爱谢小雨任常宝何玲
Owner GUANGDONG WENS DAHUANONG BIOTECH
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