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Recombinant parotitis virus particle, composition and application thereof

A mumps virus and virus technology, applied in the field of vaccines, can solve problems such as inability to effectively prevent and treat new coronavirus infection, and achieve the effect of preventing mumps virus infection, good prevention, and high poisoning rate

Pending Publication Date: 2022-04-12
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Due to the severity of the new coronavirus infection and the challenges brought by the continuous mutation of the new coronavirus to the effective prevention and / or treatment of the new coronavirus infection, the new coronavirus vaccines currently developed cannot effectively prevent and treat the new coronavirus infection

Method used

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  • Recombinant parotitis virus particle, composition and application thereof
  • Recombinant parotitis virus particle, composition and application thereof
  • Recombinant parotitis virus particle, composition and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0177] Example 1: Construction of recombinant mumps virus infectious cloning plasmids pYES2-rMuV-SC2-WH and pYES2-rMuV-SC2-Delta expressing the fusion pre-S trimer protein of the new coronavirus respectively

[0178]In this example, two recombinant mumps virus infectious cloning plasmids expressing the pre-fusion S trimer protein of the new coronavirus were constructed, pYES2-rMuV-SC2-WH and pYES2-rMuV-SC2-Delta, respectively. The construction steps of the seed plasmid are basically the same, the only difference is that the codon-optimized sequence of the S trimer protein before fusion is different. The full-length genome of the S79 strain used in the experiment (GenBank: MT732483.1) and the pre-fusion S trimer protein codon optimization of the new coronavirus prototype strain (GenBank: NC_045512.2) and the Delta strain (GenBank: OL336792.1) The sequence was synthesized by Jinweizhi Biotechnology Co., Ltd., wherein the codon-optimized sequence of the pre-fusion S trimer protei...

Embodiment 2

[0185] Example 2: Splicing of pYES2-rMuV-SC2 full-length infectious clone

[0186] Using the method of yeast transformation, the multiple fragments amplified in Example 1 were spliced ​​in yeast, specifically fragments N, P, ST1, ST2, M, F, SH-HN, L1, L2, Y1, Y2, Splicing of WH-ST1 and WH-ST2 in yeast; Fragments N, P, ST1, ST2, M, F, SH-HN, L1, L2, Y1, Y2, Delta-ST1 and Delta-ST2 in yeast stitching. Specifically: the reaction system is as follows:

[0187]

[0188] Subsequently, the yeast plasmid was extracted by a yeast plasmid extraction kit, and then electroporated into DH10B Escherichia coli competent cells.

Embodiment 3

[0189] Example 3: Verification of pYES2-rMuV-SC2-WH and pYES2-rMuV-SC2-Delta full-length infectious clones

[0190] Randomly pick individual clones, inoculate them into 5 mL of LB liquid medium containing ampicillin (100 μg / mL) resistance, culture in a shaker at 37°C for 12 h, and extract the picked clones according to the instructions of the AxyPrep Plasmid DNA Mini Kit. Then obtain the pYES2-rMuV-SC2-WH and pYES2-rMuV-SC2-Delta full-length infectious clone plasmids, perform PCR amplification on the full-length infectious clones, and verify whether the molecular weight of the PCR product meets expectations by nucleic acid electrophoresis; when the PCR product After the molecular weight is correct, the correctness of the sequence is verified by gene sequencing.

[0191] The same primers in Table 1 were used as primers for PCR sequencing, and 5 μL of PCR products were electrophoresed in 1% agarose gel for detection. detectable (results such as Figure 4 shown). Plasmids of p...

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Abstract

The invention relates to a recombinant mumps virus particle, a composition and application thereof. The recombinant mumps virus particles have the effects of high efficiency, good preventability and good safety. Therefore, the recombinant mumps virus particles and the composition comprising the same of the present invention can effectively prevent and treat new coronavirus.

Description

technical field [0001] The present invention relates to the technical field of vaccines, in particular to a recombinant mumps virus particle, a composition comprising the recombinant mumps virus particle, and the use of the composition in preparing a vaccine for preventing and / or treating new coronavirus, or the combination Use of the drug in the preparation of a drug for preventing and / or treating a disease or condition caused by a new coronavirus infection in a subject. Background technique [0002] Novel coronavirus (SARS-CoV-2) is the causative agent of novel coronavirus pneumonia (COVID-19). The new coronavirus has continuously mutated and mutated since the outbreak, and the difficulty of transmission of different mutant strains, the severity of related diseases, or the effectiveness of vaccines and therapeutic drugs are quite different. Currently, the World Health Organization has announced a new naming system for variants of the new coronavirus using the Greek alphab...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C07K14/165C12N15/50C12N15/86C12N7/04C12N15/70A61K39/295A61K39/215A61K39/165A61P31/14
Inventor 黄耀伟宗明瑞徐令东王斌杨永乐张雅晴
Owner ZHEJIANG UNIV
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