Construction and application of Zika virus ZG01 strain reverse genetic system

A Zika virus strain technology, applied in the construction field of Zika virus ZG01 strain, can solve complex and time-consuming problems

Pending Publication Date: 2020-12-01
SUN YAT SEN UNIV
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  • Abstract
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Problems solved by technology

In view of the toxicity and instability of the flavivirus genome in bacterial hosts, methods such as in vitro recombination of multi-plasmid systems and insertion of eukaryotic introns into the genome have been applied, but these methods are very complicated and time-consuming in practice

Method used

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  • Construction and application of Zika virus ZG01 strain reverse genetic system
  • Construction and application of Zika virus ZG01 strain reverse genetic system
  • Construction and application of Zika virus ZG01 strain reverse genetic system

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Embodiment Construction

[0025] The present invention will be further explained below in conjunction with specific examples, but it should be noted that the following examples are only used to explain the present invention, and can not be used to limit the present invention, and all technical solutions identical or similar to the present invention are included in the present invention within the scope of protection. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art, and the raw materials used are commercially available items.

[0026] 1) Analysis of viral sequences.

[0027] 2) Use TRIzol method to extract viral RNA from virus liquid. Take 200ul virus solution and add it to RNase-free EP tube, add 750ul TRIzol reagent, mix gently, incubate at room temperature for 10 minutes, add 200ul chloroform, shake and mix, and let stand at room temperature for 3 minutes. Centrifuge at 12,000g for 15 minutes at 4°C. Take the supern...

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Abstract

Zika virus (Zika virus, ZIKV) belongs to flaviviridae flaviviruses, and in recent years, Asian lineage viruses of Zika virus are spread to a plurality of countries in the world from South America andthe like. ZIKV infection is associated with Green-Barie syndrome and some birth defects, including forehead syndrome and the like. A reverse genetic system is an important tool for researching virus life cycle, pathogenesis, vaccine research and development and antiviral drugs, but toxicity and instability of cDNA clone of flavivirus genome in transformed bacteria are always a technical difficultyfor hindering construction of infectious clone. According to the invention, a genetically stable eukaryotic-started ZIKV full-length infectious clone ZG01 is constructed. The ZG01 reverse genetic system provides a platform and a research tool for researching a ZIKV life cycle, ZIKV-host interaction, vaccine and antiviral drug research and development. The construction method can also provide scientific reference for construction and research of reverse genetics systems of other flaviviruses or RNA viruses.

Description

technical field [0001] The invention belongs to the technical field of virus molecular biology and gene cloning, and in particular relates to the construction and application of a Zika virus ZG01 strain. Background technique [0002] Zika virus (ZIKV) belongs to the Flavivirus genus of the Flaviviridae family. In recent years, its Asian lineage virus has spread from South America and other places to many countries around the world. ZIKV infection has been linked to Guillain-Barré syndrome and some birth defects, including microcephaly. The reverse genetics system is an important tool for studying virus life cycle, pathogenesis, vaccine development and antiviral drugs, but the toxicity and instability of flavivirus genome cDNA clones in transformed bacteria have always been a major obstacle to the construction of infectious clones. Technical Difficulties. [0003] Regarding the reverse genetics system of ZIKV, in June 2016, Shan et al. first reported the construction of a f...

Claims

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Application Information

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Patent Type & AuthorityApplications(China)
IPC IPC(8): C12N7/00
CPCC12N7/00C12N2770/24121
Inventor李义平陈奕奕
OwnerSUN YAT SEN UNIV