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Seneca Valley virus recombinant plasmid and Seneca Valley virus recombinant virus, and construction methods thereof

A technology for recombining plasmids and viruses, applied in the direction of viruses, viral peptides, viruses/phages, etc., can solve the problems of SVV infection characteristics, unclear host range, and no commercial vaccines

Active Publication Date: 2018-12-18
HUAZHONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Many aspects of the infection characteristics, host range, and epidemiology of SVV are currently unknown, and there is no commercially available vaccine for prevention and control

Method used

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  • Seneca Valley virus recombinant plasmid and Seneca Valley virus recombinant virus, and construction methods thereof
  • Seneca Valley virus recombinant plasmid and Seneca Valley virus recombinant virus, and construction methods thereof
  • Seneca Valley virus recombinant plasmid and Seneca Valley virus recombinant virus, and construction methods thereof

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Experimental program
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Embodiment 1

[0040] (1) Build routes such as figure 1 shown. According to the full-length genome sequence of SVV HB-CH-2016, it was divided into four large segments: F1, F2, F3, and F4 to amplify its full-length genome. At the same time, a CMV promoter sequence from pEGFP-N1 is fused at the 5' end of the genome, and a hepatitis D virus ribozyme recognition sequence HDVr-poly sequence is fused at the 3' end. Cloning each fragment into the low-copy plasmid pBluescript II SK in sequence to obtain the infectious cloning plasmid of the SVV HB-CH-2016 strain mediated by the CMV promoter, named pSKII-CMV-SVV / HB;

[0041] (2) Rescue of SVV infectious virion (rSVV)

[0042] figure 2 For the rescue of rSVV, where, figure 2 -A is the cytopathic pattern of BHK-21 cells infected by SVV HB-CH-2016 strain and rSVV for 18 hours, figure 2 -B is the PCR identification of rSVV--using SVV-specific 5'UTR (366bp), VP3 / 1 (542bp) and 3D (298bp) genes to carry out PCR amplification of rSVV, figure 2 -C i...

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Abstract

The invention relates to a Seneca Valley virus recombinant plasmid and a Seneca Valley virus recombinant virus, and construction methods thereof, and belongs to the technical field of virus construction. The Seneca Valley virus recombinant virus is a Seneca Valley virus with the 177th amino acid, mutated to A from S, of a GH ring of capsid VP2. The Seneca Valley virus recombinant virus has a highvirus tilter and a high virus proliferation ability.

Description

technical field [0001] The invention relates to the technical field of virus construction, in particular to a Seneca Valley virus recombinant plasmid, recombinant virus and a construction method. Background technique [0002] Seneca Valley Virus (SVV), a member of picornaviruses, has oncolytic properties and can selectively infect neuroendocrine tumor cells. It has been used in preclinical research and early clinical trials Shows promise for cancer treatment. It has been reported that through genome-wide loss-of-function screening, anthrax toxin receptor 1 (ANTXR1), also known as tumor endothelial cell marker 8 (TEM8) protein, is an essential receptor for Seneca Valley virus to invade host cells. The interaction between SVV and ANTXR1 is direct and specific. This interaction requires SVV to bind to the recipient cells, and in order to avoid the antiviral activity of the interferon gene, the recipient cells need to express ANTXR1 highly, and the nucleocapsid of SVV directly ...

Claims

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Application Information

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IPC IPC(8): C12N7/01C12N15/85C12N15/66
CPCC07K14/005C12N7/00C12N15/85C12N2770/32021C12N2770/32022
Inventor 钱平李祥敏刘婷婷钱苏红陈焕春
Owner HUAZHONG AGRICULTURAL UNIVERSITY
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