System and method for rescuing measles viruses and recombinant measles viruses

A measles virus and antigenome technology, applied in the direction of virus, virus/phage, recombinant DNA technology, etc., can solve the problems of difficult to obtain stably, time-consuming and laborious, and complicated preparation process of measles vaccine, and achieves high rescue efficiency, excellent application prospect, good stability

Pending Publication Date: 2017-07-18
CHENGDU INST OF BIOLOGICAL PROD
View PDF1 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current preparation process of measles vaccine is complicated, and requires many times of passage in different cell lines to obtain attenuated strains, which is time-consuming and labor-intensive.
[0004] The patent application with publication number 103160473 discloses a method for ...

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • System and method for rescuing measles viruses and recombinant measles viruses
  • System and method for rescuing measles viruses and recombinant measles viruses
  • System and method for rescuing measles viruses and recombinant measles viruses

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Embodiment 1 The present invention rescues measles virus or constructs the recombinant measles virus vector expressing GFP

[0043] 1 Materials and methods

[0044] 1.1 Sources of vectors and strains

[0045] The measles virus Shanghai 191 originated from the Shanghai Institute of Biological Products. pT7-IRES His-C DNA was purchased from Takara Company; the vector pGEM-T was purchased from Promega Company of the United States.

[0046] 1.2 Main reagents

[0047] High-purity viral RNA extraction kit was purchased from Roche; phusion high-fidelity DNA polymerase and T4 DNA ligase were purchased from NEB; random primers, Superscript III reverse transcriptase, cell transfection reagent Lipofectamine TM 2000 was purchased from Invitrogen Company of the United States; gel recovery kit was purchased from Promega Company; plasmid extraction kit was purchased from Omega Company; agarose and DNA marker were purchased from Takara Company; other reagents were domestic reagents....

Embodiment 2

[0097] Example 2 Using the system of the present invention to rescue measles virus and recombinant measles virus expressing GFP

[0098] Rescue measles virus and recombinant measles virus expressing GFP using the vector constructed in Example 1 of the present invention: as Figure 13 As shown, build the rescue system.

[0099] 1 Rescue from the virus

[0100] BSR T7 cells were cultured in a 6-well plate at 37° C. for 24 hours, and transfected when they grew into a single layer and the confluence of the cells was 80%-90%. 12 μg full-length plasmid pT7MV S191 , 12μgpT7MV S191 -ATU-GFP was mixed with helper plasmids 8μg pT7-MV-N, 4μg pT7-MV-P, 1μg pT7-MV-L respectively, added to 100μL DMEM high-glucose culture medium and mixed, and then mixed with 100μL of premixed 10μL Lipofectamine 2000 Reagent Mix the DMEM culture solution evenly, place it at room temperature for 30 minutes, then drop it into a 6-well plate, and place it at 37°C, 5% CO 2 Cultivate in an incubator for 5-18...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses a system and a method for rescuing measles viruses and recombinant measles viruses for expressing GFP (Green Fluorescent Protein) and application. The system provided by the invention comprises the following components: 1) a full-length transcription plasmid pT7MVS191 or pT7MVS191-ATU-GFP containing a measles virus anti-genome; 2) a helper plasmid 1 containing an encoding gene of nucleocapsid protein of the measles viruses; 3) a helper plasmid 2 containing an encoding gene of phosphoprotein of the measles viruses; and 4) a helper plasmid 3 containing an encoding gene of RNA (Ribonucleic Acid) polymerase of the measles viruses. The invention further provides a method for rescuing the measles viruses and the recombinant measles viruses. By adopting the system and the method, the measles viruses and the recombinant measles viruses can be effectively rescued; and the system and the method can be used for developing and applying therapeutic or preventive recombinant measles virus products.

Description

technical field [0001] The invention belongs to a system and method for rescuing measles virus and recombinant measles virus Background technique [0002] Measles virus (measles virus) is the causative agent of measles, which belongs to the genus Measles virus in the family Paramyxoviridae. Measles is a common acute infectious disease in children. It is highly contagious and is characterized by skin papules, fever and respiratory symptoms. If there are no complications, the prognosis is good. Since the application of live attenuated vaccines in my country in the early 1960s, the incidence of children has dropped significantly. But it remains a leading cause of child mortality in developing countries. After the eradication of smallpox, the WHO has listed measles as one of the infectious diseases planned to be eliminated. Also found subacute sclerosing panencephalitis (subacute sclerosing panencephalitits, SSPE) and measles virus. [0003] MV-S191 is one of the production...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12N15/86C12N7/04
CPCC12N7/00C12N15/86C12N2760/18423C12N2760/18452
Inventor 刘兰军张勇侠李玫颖陈宗香
Owner CHENGDU INST OF BIOLOGICAL PROD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap