A kind of recombinant virus expressing classical swine fever virus e2 gene and its preparation method and application

A recombinant virus, swine fever virus technology, applied in the directions of botanical equipment and methods, biochemical equipment and methods, applications, etc., can solve the problems of drug substitution and immune clinical disease, and achieve the effect of increasing the expression amount

Active Publication Date: 2017-03-08
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the attenuated CSF vaccine can effectively prevent the infection of different gene subtypes of viruses, CSF is still prevalent in some areas, and there are clinical symptoms of virus generation and immunity

Method used

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  • A kind of recombinant virus expressing classical swine fever virus e2 gene and its preparation method and application
  • A kind of recombinant virus expressing classical swine fever virus e2 gene and its preparation method and application
  • A kind of recombinant virus expressing classical swine fever virus e2 gene and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Construction of a recombinant baculovirus Ac-CSE12 expressing the immunogenic gene E2 gene of CSFV

[0042] (1), construction of recombinant baculovirus Ac-CSE12

[0043] 1. Acquisition of the main immunogenic gene E2 of CSFV

[0044] With reference to the E2 gene sequence (GenBank No.FJ456876) of the CSFV QZ-07 isolate (2.1 type swine fever virus), the E2 gene sequence of 1222bp was artificially synthesized according to the preference of baculovirus codons, and the sequence was cloned into the vector pUC-18( Invitrogen Company) obtained the recombinant plasmid pUC-57-2.1-E2.

[0045] Using the plasmid pMD-18T-CSFV-E2 (containing the E2 gene of the rabbitized attenuated vaccine strain of classical swine fever virus HCLV, master student: Tan Huaju; supervisor: Qian Ping; Huazhong Agricultural University 2009 master's thesis: BVDV E2 gene recombinant classical swine fever virus Construction and expression of porcine IRF3 / 7 clones) as templates, using primers 1.1E2-GZF a...

Embodiment 2

[0071] Subunit vaccine prepared by recombinant baculovirus Ac-CSE12 of the present invention and immune efficacy experiment

[0072] 1. The subunit vaccine prepared by the recombinant baculovirus Ac-CSE12 of the present invention

[0073] The recombinant baculovirus Ac-CSE12 obtained in Example 1 was used to inoculate the suspension-cultured insect cells High Five at a MOI dose of 1.0 TM (purchased from Invitrogen Company), after 72 hours, the cells and supernatant were collected, processed by a cell disruptor, centrifuged to remove cell debris, and after measuring the protein concentration, it was emulsified with 206 adjuvant (purchased from SEPPIC Company) to prepare a sub-vaccine. The protein content of CSFV E2 in the medium is 40 micrograms, used in the following examples, and stored at 4°C.

[0074] 2. Inspection of subunit vaccine technical indicators prepared by recombinant baculovirus Ac-CSE12 of the present invention

[0075] (1) Physical shape

[0076] Appearance ...

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Abstract

The invention discloses a recombinant virus for expressing swine fever virus E2 gene, and a preparation method and application thereof. According to the preference of the codon, synthetic CSFV 2.1-type immunogenic gene E2 and 1.1-type E2 gene are respectively arranged in baculovirus pH and p10 promoters to be expressed so as to obtain a recombinant baculovirus (CCTCC NO:V201338). The exogenous gene expression level is enhanced. After being used for immunizing Japanese white rabbits and pigs, the subunit vaccine prepared by the E2 protein expressed by the recombinant baculovirus can induce the organism to generate specific immune reaction and protect the organism from the attack of the swine fever virus; and differential diagnoses can be utilized to distinguish the vaccine immunized animal from the wild virus-infected animal, thereby being beneficial to eradication and purification of the eradicate swine fever virus in the pig farm.

Description

technical field [0001] The invention belongs to the field of veterinary biotechnology. The invention relates to a method for constructing a recombinant baculovirus strain. In particular, it relates to a preparation method of recombinant baculovirus Ac-CSE12 expressing classical swine fever virus E2 gene and its application in a subunit vaccine for preventing classical swine fever infection. The artificially modified CSFV E2 gene expressed by the recombinant virus of the invention can be used to prepare a safe CSFV subunit vaccine. Background technique [0002] Classical swine fever (CSF) is a highly contagious, hemorrhagic and fatal infectious disease caused by Classical swine fever virus (CSFV) of the Flaviviridae family. The World Organization for Animal Health (OIE) lists it as an animal disease that must be reported, and my country lists it as a first-class animal disease. At present, swine fever is prevalent in many pig farms in my country, clinically manifested as p...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/62C12N15/866C12N15/66C12N7/01C07K19/00A61K39/12A61P31/14G01N33/68
Inventor 李祥敏钱平张华伟朱世轩钱苏红陈焕春
Owner HUAZHONG AGRI UNIV
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