Mutant foot-and-mouth disease virus infectious clone and preparation method and application thereof

A technology for foot-and-mouth disease virus and infectious cloning, which is applied in the field of viruses in biotechnology, and can solve the problems of SUMO modification that have not been reported.

Active Publication Date: 2020-02-14
INST OF ANIMAL SCI & VETERINARY MEDICINE SHANDONG ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there is no report on the role of SUMOylation modification in the process of FMDV infection

Method used

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  • Mutant foot-and-mouth disease virus infectious clone and preparation method and application thereof
  • Mutant foot-and-mouth disease virus infectious clone and preparation method and application thereof
  • Mutant foot-and-mouth disease virus infectious clone and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] 1. Construction of FMDV IC-K5154110159R infectious clone

[0031] 1.1 Primer design

[0032] According to the O-type FMDV Tibet / CHA / 99 strain (GenBank accession number: AJ539138) sequence design primers for amplifying the three fragment sequences of A, B, and C, as follows:

[0033] figure 1 A, B, C fragment amplification primers

[0034]

[0035] Restriction sites are underlined; T7 promoter is shown in bold

[0036] 1.2 Construction and identification of full-length cDNA plasmid containing O-type FMDV Tibet / CHA / 99 strain

[0037] (1) Extract viral total RNA with TRIzol reagent, use reverse transcriptase SuperScript of Invitrogen Company TM III Reverse Transcriptase performed reverse transcription according to the instructions, and then used the high-fidelity enzyme PfuUltra TM II Fusion HS DNA Polymeras and the designed primers FMDV-A-Fwd / Rev, FMDV-B-Fwd / Revand FMDV-C-Fwd / Rev PCR amplified the three fragments A, B, and C, and the results are shown in figure...

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Abstract

The invention relates to the technical field of viruses in biotechnology, in particular to a mutant foot-and-mouth disease virus infectious clone, and also relates to preparation and application of the infectious clone. Lysine at the 51st, 54th, 110th and 159th sites of 3C protein is mutated into arginine through a site-specific mutagenesis technology. According to the constructed mutant infectious clone FMDV IC-K5154110159R, SUMOylation modification sites of the 3C protein of the infectious clone are mutated, the infectious clone can be proliferated, and a powerful tool is provided for detecting and researching FMDV infection and SUMOylation modification at the virus level.

Description

technical field [0001] The invention relates to the technical field of viruses in biotechnology, in particular to a mutant foot-and-mouth disease virus infectious clone, and also relates to the preparation and application of the infectious clone. Background technique [0002] Foot-and-mouse disease (FMD) is an acute, febrile, highly contagious disease of cloven-hoofed animals caused by foot-and-mouse disease virus (FMDV). The outbreak of the disease has caused great losses to animal husbandry production and people's lives, and it has been listed as the first class A animal infectious disease by the International Veterinary Bureau. FMDV belongs to the Picornaviridae family and the genus Foot-and-Mouth Disease Virus. The genome is a single-stranded positive-sense RNA with a total length of about 8500bp. It consists of a 5' non-coding region (UTR), a large open reading frame (ORF) and a 3' non-coding Region (UTR) composition. The ORF encodes a large polyprotein that is cleave...

Claims

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Application Information

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IPC IPC(8): C12N15/85C12N7/00C07K14/09
CPCC07K14/005C12N7/00C12N15/85C12N2770/32121C12N2770/32122C12N2770/32152C12N2800/107
Inventor 吴香菊杜以军齐静丛晓燕隋超
Owner INST OF ANIMAL SCI & VETERINARY MEDICINE SHANDONG ACADEMY OF AGRI SCI
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