Construction of recombinant rabies virus of double-expression G gene and biological property analysis thereof

A rabies virus, recombinant virus technology, applied in microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve problems such as mouse infection, and achieve immunogenic effects

Inactive Publication Date: 2010-07-07
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The virus is not pathogenic in dogs but retains its immunogenicity, but remains infective and fatal in mice wh...

Method used

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  • Construction of recombinant rabies virus of double-expression G gene and biological property analysis thereof
  • Construction of recombinant rabies virus of double-expression G gene and biological property analysis thereof
  • Construction of recombinant rabies virus of double-expression G gene and biological property analysis thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1 Contains the construction and identification of the recombinant virus of two G genes

[0026] 1 Materials and methods

[0027] 1.1 Plasmids, cell lines and viruses

[0028] Plasmids pCAGG and pCI-RBz were purchased from Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, and pBluescriptSK(+) was purchased from clonetech. BHK-21 cells (ATCC CC1-10) and 293T cells (CRL-11268) were purchased from ATCC, and the culture medium was DMEM (purchased from GIBCO) containing 10% fetal bovine serum. Mouse brain neuroma cells NA cells come from the Military Veterinary Research Institute of the Academy of Military Medical Sciences, and the culture medium is MEM containing 10% fetal bovine serum (purchased from GIBCO). Rabies virus Flury-LEP and HEP were purchased from the China Veterinary Microbial Culture Collection Management Center of the China Veterinary Drug Administration, amplified on BHK-21 cells, and frozen at -70°C for future use. ...

Embodiment 2

[0083] The immunization test of embodiment 2 recombinant virus Beagles

[0084] In order to evaluate the immune effect of the recombinant virus on dogs, 8×10 7 FFU / ml of wtLEP and rLEP-G were treated with 1:6000β-propiolactone for 2h to inactivate, mixed with aluminum phosphate adjuvant (purchased from REHEIS company) 3:1, and immunized by intramuscular injection for 3 months. 6 greyhounds (purchased from the Experimental Animal Center of the Academy of Military Medical Sciences of the Chinese People's Liberation Army) each, and 6 dogs were used as controls, and 1ml of PBS was injected intramuscularly. Blood was collected 21 days after immunization and serum was separated for detection of neutralizing antibody titers. Neutralization test

[0085] Serum samples were first inactivated by treatment in a water bath at 56°C for 30 minutes, and then serially diluted from 20 times to 1280 times. At the same time, negative and positive serum controls were set. The standard immobili...

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Abstract

The invention provides a recombinant rabies virus strain with a G gene additionally inserted in a genome, a method for constructing the recombinant rabies virus strain and application thereof. The recombinant rabies virus strain constructed by the method has high safety and immunogenicity, and is suitable for preparing vaccines for inactivating rabies virus.

Description

Technical field: [0001] The invention belongs to the field of recombinant vaccines. Specifically, the present invention provides a recombinant rabies virus strain, which additionally inserts a G gene into the genome; the present invention provides a method for constructing the recombinant rabies virus strain and its application for preparing a vaccine. Background technique [0002] Rabies is an important zoonotic disease responsible for approximately 60,000 deaths per year (1) . In many developing countries, dogs are still the main cause of human rabies, while in developed countries, rabies virus is mainly found in wild animal reservoirs. Practice has proved that immunization of animals such as dogs can significantly reduce the incidence of human rabies (2) , so immunization of animals is extremely important for the control of rabies. [0003] Rabies virus belongs to the genus Lyssavirus in Rhabdoviridae. The genome is a single-stranded negative-sense RNA without segment...

Claims

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Application Information

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IPC IPC(8): C12N7/01C12N15/47C12N15/85A61K39/205A61P31/14C12R1/93
Inventor 步志高陶丽红葛金英王喜军
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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