System used for rescuing mumps virus and rescue method

A technology of mumps virus and helper plasmids, applied in the field of reverse genetics, which can solve problems such as complex systems and methods, difficult rescue, etc., and achieve the effect of shortening time and cost, and overcoming time-consuming and labor-intensive effects

Pending Publication Date: 2019-04-09
ZHEJIANG UNIV
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AI Technical Summary

Benefits of technology

This patented technology allows researchers to efficiently recover viruses that cause outbreaks or diseases like measles disease through genetic engineering techniques such as inserting foreign DNA into infected host cell genes during their replication cycle instead of producing them separately on an expensive scale. Additionally, this new method enables quicker testing and create small amounts of material needed for making effective immunotherapies against these types of illnesses.

Problems solved by technology

Technological Problem addressed in this patents relates to developing safe and efficient ways to protect against severe illness caused by rhinorrhinosus coronavirus (RSV)-related conditions like pneumonia or influenza. Current measures involve administering antiinfluenzolvals drugs with limited effectiveness due to their side effects. There exists a technical problem related to finding simpler and easier way to prepare these medicaments while maintaining efficacy over longer periods.

Method used

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  • System used for rescuing mumps virus and rescue method
  • System used for rescuing mumps virus and rescue method
  • System used for rescuing mumps virus and rescue method

Examples

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Embodiment 1

[0052] 1. Extract viral genome RNA

[0053] The mumps S79 attenuated vaccine strain was provided by the Zhejiang Provincial Center for Disease Control and Prevention, according to TRIzol TM The Reagent cell lysate instructions were used to extract the genomic RNA of the vaccine strain.

[0054] 2. PCR amplification to prepare DNA fragments of the whole genome of the S79 vaccine strain

[0055] Using the extracted mumps virus RNA as a template, according to III Reverse transcription kit instructions, use Random Primer Mix to reverse-transcribe mumps virus RNA into cDNA, and store at minus 20°C for later use.

[0056] There are 8 DNA fragments in the full-length genome of the S79 vaccine strain, and the 8 fragments are NP, P, M, F, SH, HN, L1 and L2 respectively. Using cDNA as a template, 5 pairs of overlapping primers are used to amplify and prepare Covering the full-length genome of the S79 vaccine strain, the primer sequences are shown in Table 1. The original fragment ...

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Abstract

The invention discloses a system used for rescuing a mumps virus and a rescue method, and belongs to the field of application of reverse genetics. The system comprises a recombinant transcription vector containing a whole genome cDNA (complementary deoxyribonucleic acid) of the mumps virus, a helper plasmid I containing an NP gene of the mumps virus, a helper plasmid II containing a P gene of themumps virus and a helper plasmid III containing an L gene of the mumps virus. The mumps virus can be effectively rescued from outside of a body by adopting a 3+1 plasmid rescue system (helper plasmidsand the transcription vector that are used for expressing an N protein, a P protein and an L protein of a MuV-S79 strain), the recombinant mumps virus can be abundantly amplified by a cell culture method, and the recombinant mumps virus can be directly used for manufacturing vaccines without any processing, so that the problems of the traditional vaccine that the production is time-saving and labor-saving, and the process is complex are solved. Additionally, a solution is provided for developing a novel vaccine by engineering the mumps virus by adopting a molecular clone technology in the future.

Description

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Claims

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Application Information

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Owner ZHEJIANG UNIV
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