Porcine BVDV (Bovine Viral Diarrhea Virus) recombinant virus for expressing porcine epidemic diarrhea virus (PEDV) S genes and application

A porcine epidemic diarrhea and recombinant virus technology, applied in the direction of virus, virus/phage, recombinant DNA technology, etc., can solve the problems that have not been reported, the vaccine cannot provide immune protection, and the economic loss of the pig industry.

Inactive Publication Date: 2018-10-02
SHANGHAI ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0005] In 2011, porcine epidemic diarrhea was prevalent in my country on a large scale, causing serious economic losses to the pig industry. The mutation of PEDV strains, especially the mutation of the S gene, and the inability of existing vaccines to provide sufficient immune protection were the reasons for the epidemic. The main reason
At present, there is no report on the combination of PEDV and BVDV

Method used

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  • Porcine BVDV (Bovine Viral Diarrhea Virus) recombinant virus for expressing porcine epidemic diarrhea virus (PEDV) S genes and application
  • Porcine BVDV (Bovine Viral Diarrhea Virus) recombinant virus for expressing porcine epidemic diarrhea virus (PEDV) S genes and application
  • Porcine BVDV (Bovine Viral Diarrhea Virus) recombinant virus for expressing porcine epidemic diarrhea virus (PEDV) S genes and application

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Embodiment Construction

[0042] The present invention will be further described below in conjunction with specific examples.

[0043] DL15,000 DNA marker, pfu polymerase, Trizol, Reverse Transcriptase M-MLV(RNaseH-), 5×Reverse Transcriptase M-MLV Buffer, Recominant Rnase Inhibitor, dNTPMixture (2.5mM each), and restriction enzymes were purchased from TaKaRa Company; DMEM medium and Opti-MEM medium were purchased from Gibco; Spin Miniprep Kit was purchased from QIAGEN Company; MEGAscript T7 was purchased from Ambion Company; liposome Lipofectamine 2000 was purchased from Invitrogen Company; FITC-labeled goat anti-mouse IgG was purchased from Boster Company; other conventional reagents were domestic analytical grade products. The porcine BVDV-2 infectious cloning vector plasmid pASH28 was obtained from the Laboratory of Shanghai Academy of Agricultural Sciences.

[0044] The construction and application of embodiment porcine source BVDV-2 recombinant infectious cloning plasmid

[0045] 1. Primer Desi...

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Abstract

The invention discloses a porcine BVDV (Bovine Viral Diarrhea Virus) recombinant virus for expressing porcine epidemic diarrhea virus (PEDV) S genes and the application thereof. A neutralization antigenic epitope sequence of the PEDV S genes is inserted into a terminal N of a C nucleocapsid protein gene of porcine BVDV. A recombinant infectious cloning plasmid is obtained. The plasmid is linearized, subjected to in vitro transcription to obtain RNA and transfected with MDBK (Madin-Darby Bovine Kidney) cells, so that the porcine BVDV recombinant virus for expressing the PEDV S proteins can be successfully rescued. The porcine BVDV recombinant virus is capable of expressing a PEDV S foreign protein, and is simultaneously used for controlling the PEDV and BVDV. The porcine BVDV infectious clone provides suitable insertion sites for other exogenous genes, can serve as a recombinant viral vector, and provides an effective technology platform for researches such as subsequent in-vitro modification of RNA, insertion of exogenous genes, preparation of recombinant viruses and the like.

Description

technical field [0001] The invention belongs to the technical field of veterinary biological products, in particular to a porcine BVDV recombinant virus expressing porcine epidemic diarrhea virus S gene and application thereof. Background technique [0002] Bovine viral diarrhea virus (BVDV) and classical swine fever virus (CSFV) belong to the genus Pestivirus of the family Flaviviridae, and have certain cross-reactivity in serology. [0003] In addition to infecting cattle, BVDV can also infect pigs. The clinical symptoms or latent infection caused by it can affect all aspects of pig production, such as growth retardation, reproductive disorders, secondary infection and persistent infection. The disease is prevalent in almost all provinces and cities in China. Deng Yu et al. used nested RT-PCR to screen clinical samples in some areas of China from 2007 to 2010, and the positive rate of BVDV was 23.1% (2007). ), 27.7% (2008), 33.6% (2009) and 23.6% (2010). Pigs infected wi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/01C12N15/85C12R1/93
CPCC12N7/00C12N15/85C12N2770/20022C12N2770/24321C12N2770/24352
Inventor 刘惠莉陶洁李本强程靖华石迎饶柏钟
Owner SHANGHAI ACAD OF AGRI SCI
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