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System and method for rescuing mumps virus

A mumps virus, rescue system technology, applied in the field of virus reverse genetics, can solve problems such as labor and time consumption

Pending Publication Date: 2020-06-02
CHENGDU INST OF BIOLOGICAL PROD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Purification of JL1 substrains by limiting dilution requires serial dilution to single clones, and sequencing of the diluted single clones requires a lot of manpower and time.

Method used

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 Mumps virus rescue system construction

[0039] 1 Materials and methods

[0040] 1.1 Cells and plasmids

[0041] P139 generation Vero cells, mumps vaccine strain S 79 , plasmid pT7-IRES His-CDNA (Takara), and plasmid pCDIBP-T7RNAP. (with CMV promoter sequence and T7RNA polymerase DNA sequence, see Appendix 1, SEQ ID NO.2)

[0042] 1.2 Main reagents

[0043] High-purity viral RNA extraction kit was purchased from Roche; High-fidelity DNA polymerase, T4 DNA ligase, restriction endonuclease, etc. were purchased from NEB Company; random primers and SuperscriptIII reverse transcriptase were purchased from Invitrogen Company; gel recovery kit was purchased from Promega Company; plasmid extraction kit was purchased from Omega Company; agarose and DNAmarker were purchased from Takara Company; MEM and FBS culture medium were purchased from Gibco Company; other reagents were domestic reagents.

[0044] 1.3 Construction of full-length plasmid and helper plasmid

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Abstract

The invention provides a system and method for preparing a mumps virus. A mumps virus full-length plasmid having a T7 promoter and auxiliary plasmids are constructed, and the mumps virus is prepared by a reverse genetics method, wherein the four auxiliary plasmids are as follows: a NP plasmid, a P plasmid, an L plasmid and a T7RNAP plasmid, and the T7RNAP plasmid has the sequence of a CMV promoterand the sequence of a DNA for encoding T7 polymerase. The system and method can effectively prepare the mumps virus with higher immunogenicity, and have good application prospects.

Description

technical field [0001] The invention relates to the field of virus reverse genetics, in particular to a system and method for rescuing mumps virus. Background technique [0002] Mumps virus (MuV) belongs to the family Paramyxoviridae and belongs to the genus Mumps virus. The genome is a single-strand negative-strand ribonucleic acid (RNA) with a length of 15384bp. Mumps (mumps) is an acute infectious disease caused by the mumps virus, which can widely involve various organs, and its clinical manifestations are mainly in the parotid gland, gonads and central nervous system, such as aseptic meningitis and encephalitis , orchitis, oophoritis, pancreatitis, etc. [1] . [0003] Mumps vaccination has a history of nearly 50 years. The live attenuated mumps vaccine strains used or used at home and abroad include: Chinese S 79 strain, American Jeryl Lynn strain, Japanese Urabe Am9 strain, former Soviet Union Leningrad-Zagreb strain, Swiss Rubini strain and Belgian RIT4385 strain ...

Claims

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Application Information

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IPC IPC(8): C12N15/85C12N15/45C12N7/00
CPCC12N15/85C12N7/00C12N2760/18752
Inventor 刘兰军张勇侠高雅丽陈宗香康庄
Owner CHENGDU INST OF BIOLOGICAL PROD
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