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Marked Pestivirus suis C strain expressing enhanced green fluorescent protein and construction method and application thereof

A technology of green fluorescent protein and swine fever virus, applied in the direction of viruses, viral peptides, antiviral agents, etc., can solve the problem of inability to identify infected animals and immune animals

Inactive Publication Date: 2016-08-17
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, as an excellent vaccine strain with independent intellectual property rights in China, it has the disadvantage of being unable to distinguish infected animals from immune animals serologically

Method used

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  • Marked Pestivirus suis C strain expressing enhanced green fluorescent protein and construction method and application thereof
  • Marked Pestivirus suis C strain expressing enhanced green fluorescent protein and construction method and application thereof
  • Marked Pestivirus suis C strain expressing enhanced green fluorescent protein and construction method and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Construction of labeled CSFV strain C expressing enhanced green fluorescent protein and its biological characteristics and immunity evaluation

[0037] 1. Materials and methods

[0038] 1.1 Plasmids, cells and viruses

[0039]The infectious clone pOKHCLV of the C strain previously constructed by the inventor's laboratory (Wang Mingjie. Construction of cDNA molecular cloning and chimeric molecular cloning of hog fever lathe attenuated vaccine strain. Xinjiang Agricultural University, 2006.) and the 5' of pSM-HCLV5NCR The half-end and the 3′ half-end of pSM-HCLV3NCR (Li Chao. Molecular basis for the adaptation of a lanic attenuated vaccine strain of classical swine fever to rabbits. Chinese Academy of Agricultural Sciences, 2013; Li, C., Li, Y., Shen, L., Huang, J.,Sun,Y.,Luo,Y.,Zhao,B.,Wang,C.,Yuan,J.,Qiu,H.J.The role of noncoding regions of classical swine fever virus C-strain in its adaptation to the rabbit .Virus Res.2014,183:117-122.) was prepared and pre...

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Abstract

The present invention discloses a marked Pestivirus suis C strain expressing enhanced green fluorescent protein and construction method and application thereof, and belongs to construction and application of marked Pestivirus suis C strain. A C strain infectious clone is used as a skeleton, the Pestivirus suis Shimen strain Npro protein is used to substitute a C strain Npro protein, then an EGFP gene is introduced between No. 13 and 14 amino acids of the Npro protein to obtain an marked Pestivirus suis C strain expressing enhanced green fluorescent protein, wherein the strain has microbial preservation number: CGMCC No. 12039. The marked Pestivirus suis C strain can stably express EGFP gene, is successfully observed with green fluorescence, retains biological properties and immunogenicity of the parental viruses, and can be applied to the preparation of marked vaccines for prevention of Pestivirus suis. The present invention confirms that the C strain Npro protein can be modified to obtain marked Pestivirus suis, so as to lay foundation for the development of marked C strain vaccine.

Description

technical field [0001] The present invention relates to a labeled CSFV C strain, in particular to a labeled CSFV C strain expressing enhanced green fluorescent protein, and to a method for constructing a labeled CSFV C strain expressing enhanced green fluorescent protein and its preparation The application in marked vaccines belongs to the field of construction and application of marked classical swine fever virus C strain. Background technique [0002] Classical swine fever (CSF) is a highly contagious infectious disease of pigs characterized by high fever retention and extensive bleeding. It is listed as a Notifiable zoonotic disease by the World Organization for Animal Health (OIE). China's "National Medium and Long-Term Animal Disease Prevention and Control Plan (2012-2020)" lists it as one of the five priority animal diseases. [0003] Classical swine fever virus (CSFV) is the pathogen of CSF, which belongs to the member of the genus Pestivirus in the family Flavivirid...

Claims

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Application Information

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IPC IPC(8): C12N7/01C12N15/86C12N15/62A61K39/187A61P31/14
CPCC12N7/00A61K39/187C07K14/18C07K2319/60C12N2770/24334
Inventor 仇华吉李永锋王晓张玲楷孙元李素罗玉子
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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