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Novel recombinant coronavirus based on vesicular stomatitis virus vector as well as preparation method and application of novel recombinant coronavirus

A technology of vesicular stomatitis and recombinant virus vector, which is applied in the field of biomedicine, can solve the problems of lack of targeted specific drugs and treatment methods, and achieve the effect of easy preparation, easy cultivation, and good safety

Active Publication Date: 2021-06-11
INST OF ZOOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the new type of coronavirus 2019-nCoV (SARS-CoV-2) is in the stage of cumulative understanding, and patients are mainly given symptomatic treatment and supportive treatment, lacking targeted specific drugs and treatment methods

Method used

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  • Novel recombinant coronavirus based on vesicular stomatitis virus vector as well as preparation method and application of novel recombinant coronavirus
  • Novel recombinant coronavirus based on vesicular stomatitis virus vector as well as preparation method and application of novel recombinant coronavirus
  • Novel recombinant coronavirus based on vesicular stomatitis virus vector as well as preparation method and application of novel recombinant coronavirus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0082] Example 1 Preparation of recombinant virus RVSV-2019-NCOV (RVSV-SARS-COV-2)

[0083] I. Preparation of recombinant vector VSV-2019-NCOV

[0084] 1. Optimization of the vesic protein S sequence

[0085] In order to develop the 2019-NCOV vaccine, the sequence encoding the complete 2019-NCOV capsule protein S is subjected to humanized optimization, and the optimized sequence is shown in Sequence 1 in the sequence list. The complete 2019-NCOV capsule protein S is a 2019-NCOV Wuhan-HU-1 strain (GenBank: YP_045512.2) vesicular protein S (GenBank: YP_009724390.1), and its amino acid sequence is in a sequence list Sequence 2 is shown.

[0086] 2, restructuring vector VSV-2019-NCOV construction

[0087] The optimized sequence (sequence 1 in the sequence list) in step 1 is inserted between the enzyme digestion of the RVSVΔg vector, and the recombinant carrier is obtained, and it is named recombinant vector Vsv-2019-NCOV.

[0088] Second, the preparation of recombinant virus RVSV-2019...

Embodiment 2

[0093] Example 2, Preparation of recombinant virus RVSV-2019-NCOV-SARS

[0094] First, the preparation of recombinant vector VSV-2019-NCOV-SARS

[0095] 1, optimization of chimeric S sequence

[0096] In order to develop a 2019-NCOV vaccine, a sequence encoding the sacral protein S made from the extracellular region of 2019-NCOV velocytoprotein s extracellular zone and the sac sac sacral region and the intracellular region of the intracellular region. Source-optimization, the optimized sequence is shown in Sequence 3 in the sequence list. The 2019-NCOV vesicular protein S extracellular region is the No. 2019-NCOV Wuhan-Hu-1 strain of the venom protein S extracellular region (the amino acid sequence of the bladder protein S of the Wuhan-HU-1 strain 1-1210 bits), SARS-COV vesicular protein S transmembrane region and intracellular area are SARS-COV (Severe Acute Respiratory Syndrome-Coronavirus) BJ01 strain (GenBank: AY278488.2) of the vesicle protein S transmembrane Zone and intrace...

Embodiment 3

[0101] Example 3, Preparation of recombinant virus RVSV-2019-NCOV '

[0102] First, the preparation of recombinant vector VSV-2019-NCOV '

[0103] 1. Optimization of the vesic protein S sequence

[0104] In order to develop the 2019-NCOV vaccine, a sequence encoding a saccharin S made from a saccharin G transmembrane region of 2019-NCOV capsule protein S progefa and a VSV chunted cytosome protein S is subjected to humanized optimization. Optimized sequences shown in sequence 5 in the sequence list. The 2019-NCOV vesicular protein S extracellular region is the extracellular region of the 2019-NCOV Wuhan-HU-1 strain of the venoms. The VSV glycoprotein G transmembrane region and intracellular region are glycoprotein G transmembrane regions and intracellular region of VSV Indiana strains. The amino acid sequence of the saccharin S formed by a glycoprotein g of 2019-NCOV, a glycoprotein g of a VSV, a saccharin S, is shown in Sequence 6 in the sequence list in the sequence list.

[0105...

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Abstract

The invention discloses a novel recombinant coronavirus based on a vesicular stomatitis virus vector as well as a preparation method and application of the novel recombinant coronavirus. The novel recombinant coronavirus is a virus obtained by replacing glycoprotein G of vesicular stomatitis virus with envelope protein S; the envelope protein S comprises an envelope protein S extracellular region of 2019-nCoV (SAR-CoV-2) or a part of the sequence of the envelope protein S extracellular region. Experiments prove that the recombinant novel coronavirus can simulate the process that 2019-nCoV (SARS-CoV-2) invades cells, and can stimulate an organism to generate immunoreaction aiming at the 2019-nCoV (SARS-CoV-2), so that the recombinant novel coronavirus is an ideal tool for researching the infection process of the 2019-nCoV (SARS-CoV-2), can be developed into vaccines, and has a good application prospect. the recombinant novel coronavirus has important significance for screening inhibitors for inhibiting virus invasion and developing vaccines.

Description

Technical field [0001] The present invention relates to the field of biopharmaceutical technology, and in particular to replica restructuring new coronaviruses based on blisteritis virus vectors and their preparation methods and applications. Background technique [0002] The new coronavirus 2019-NCOV is also called SARS-COV-2 or HCOV-19, which is a new β-crowned virus. Viral genome analysis showed that the virus was very close to Sars-CoV (Severe Acute Respiratory Syndrome-Coronavirus) virus (89.1% nucleotide similarity), which caused symptoms such as human pneumonia after infection with new coronaviruses 2019-NCOV. The new coronavirus 2019-NCOV (SARS-COV-2) belongs to a single stranded RNA virus with a capsule. It is observed under electron microscopy, the viral particles are circular or elliptical, with an emitted protrusion. Such an emissive projection is a characteristic structure of a coronavirus which is composed of a saccharin spike (s). S proteins are an important protei...

Claims

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Application Information

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IPC IPC(8): C12N7/01C12N15/85C12N15/65C12N15/50C12N15/62A61K39/215A61P31/14C12R1/93
CPCA61K39/12A61K2039/5256A61P31/14C07K14/005C07K2319/00C12N7/00C12N15/65C12N15/85C12N2760/20221C12N2760/20222C12N2760/20252C12N2770/20022C12N2770/20034C12N2800/107
Inventor 郑爱华赵超越李虹悦张毓航金万洙赵建国
Owner INST OF ZOOLOGY CHINESE ACAD OF SCI
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