Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Method for constructing adenovirus for efficient package and expression of melanoma differentiation-associated gene-7

A melanoma and construction method technology, which is applied in the field of adenovirus construction for efficient packaging and expression of melanoma differentiation-related gene-7, can solve the problems of low recombination efficiency, complicated operation, and poor anti-tumor selectivity

Inactive Publication Date: 2015-07-08
张雅洁 +1
View PDF2 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide an adenovirus construction method for efficiently packaging and expressing melanoma differentiation-related gene-7, which overcomes the defects of complicated operation, low recombination efficiency, and poor anti-tumor selectivity in the prior art adenovirus construction method

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
  • Method for constructing adenovirus for efficient package and expression of melanoma differentiation-associated gene-7
  • Method for constructing adenovirus for efficient package and expression of melanoma differentiation-associated gene-7
  • Method for constructing adenovirus for efficient package and expression of melanoma differentiation-associated gene-7

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0020] The present invention is mainly divided into two steps. The first step is the construction of the adenovirus expression vector, that is, the pDC316 belonging to the adenovirus AdMax is used as the shuttle vector to construct the recombinant adenovirus expression vector instead of the most popular AdEasy system. The main method is: co-transfect the recombinant shuttle plasmid vector pDC316-hMDA7 containing the target gene hMDA-7 fragment and the backbone plasmid pBHGloxΔE1,3Cre into HEK293 cells, package and produce recombinant adenovirus Ad.mda-7.egfp, and Amplification and infectious titer determination were performed. Step 1 is mainly to ensure the success rate of the AdMax system, that is, to ensure that the AdMax system recombines the eukaryotic cells to produce the virus, and the success rate of the virus is greater than 98%. The specific operation steps are as follows.

[0021] 1. Packaging of recombinant adenovirus Ad.mda-7.egfp.

[0022] The adenoviral vector p...

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 provides a method for constructing an adenovirus for efficient package and expression of melanoma differentiation-associated gene-7 (mda-7). The method adopts pDC316 belonging to adenovirus AdMax<TM> as a vector to construct and recombine an adenovirus expression vector to replace a most popular AdEasy system. Compared with the present most popular adenovirus AdEasy system, an adenovirus AdMax<TM> system has the characteristics of being simple and convenient to operate, high in recombination efficiency and high in yield (generally larger than 10<4>vp / cell) of obtained viruses. The method adopts the mda-7 which selectively induces apoptosis of tumor cells as a therapeutic gene to construct a replication-defective adenovirus vector Ad.mda-7.egfp through package of the mda-7 gene via the AdMax<TM> system, and testifies that the replication-defective adenovirus vector Ad.mda-7.egfp has the function of selectively inducing apoptosis of lung adenocarcinoma cell line A549, thereby laying a favorable experimental foundation for gene therapy of lung adenocarcinoma.

Description

technical field [0001] The invention relates to a virus construction method, in particular to an adenovirus construction method for efficiently packaging and expressing melanoma differentiation-related gene-7. Background technique [0002] Common packaging methods for adenoviral vectors include: 1) Classic homologous recombination method (two-plasmid co-transfection): The principle is homologous recombination. Insert the target gene expression cassette into the left arm region of the adenovirus (usually missing the E1 gene region) to form an adenovirus shuttle plasmid, and co-transfect cells carrying the E1 gene region with the large plasmid containing the entire genome of the adenovirus (usually missing the E1 gene region) Like 293 cells, the two plasmids form a recombinant adenovirus genome through homologous recombination in 293 cells, and are packaged into virus particles. 2) Ad-Easy method: the principle is to obtain recombinant adenovirus genome plasmids in bacteria t...

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): C12N15/861C12N15/66
Inventor 张雅洁李书华
Owner 张雅洁
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com