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Construction method and application of an improved adenovirus and its system

A construction method and technology of adenovirus, applied in the field of adenovirus, can solve the problems of false positive virus affecting immunity and treatment effect, and achieve the effect of eliminating the effect of false positive

Active Publication Date: 2021-11-05
SHENZHEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The object of the present invention is to provide a construction method and application of an improved adenovirus and its system, which solves the problem that the virus particles in the adenovirus system in the prior art express fiber protein and the fusion protein of fiber and target protein mixed virus particles lead to a large number of False-positive viruses that only express fiber protein affect immunity and therapeutic effects

Method used

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  • Construction method and application of an improved adenovirus and its system
  • Construction method and application of an improved adenovirus and its system
  • Construction method and application of an improved adenovirus and its system

Examples

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Embodiment

[0042] Example: Taking the preparation of human defensin hFIZZ3 monoclonal antibody as an example, the above-mentioned construction method of the present invention and related experimental descriptions are explained in detail

[0043] Design primers to amplify hFIZZ3 (about 300bp) from human cDNA, introduce NotI and XhoI at both ends of the gene, the primer sequence is: upstream primer Not I-hf3-F: 5'-TCTGCGGCCGCAG ATG AAG CGC GCA AGA CCG TCT-3' ; Downstream primer XhoI-hf3-R: 5'-GGC CTCGAG GGG CTG CAC ACG ACA GCA GCG CGC-3', digested and loaded into the transformed vector pShuttle-CMV-fiber to obtain pShuttle-CMV-fiber-hFIZZ3 After linearization, it was recombined with BJ-5183-AD bacteria containing the virus backbone, and the monoclonal recombinant was cultured to extract plasmids for enzyme digestion and identification, and the positive plasmid pShuttle-cmv-fiber-hFIZZ3-AD was obtained, and pShuttle- cmv-fiber-hFIZZ3-AD plasmid, recover a 30kbp DNA fragment, transfect the 3...

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Abstract

The present invention relates to a construction method and application of an improved adenovirus and its system. After the fiber gene is knocked out in AD293 cells, the surface of the adenovirus particle no longer only expresses fiber protein, but expresses a fusion protein of fiber and a target protein. On the surface of virus particles, there are only fusion proteins of fiber and target protein, which eliminates the false positive effect of virus particles with only fiber protein.

Description

technical field [0001] The present invention relates to the technical field of adenovirus, more specifically, relates to a construction method and application of an improved adenovirus and its system. Background technique [0002] Viral vectors are widely used in immunization and gene therapy, among which the adenovirus system is currently the most widely used tool. The virus particles in the existing adenovirus system are mixed virus particles expressing fiber protein and fusion protein of fiber and target protein, but a large number of false positive viruses expressing only fiber protein will affect the effect of immunity and treatment. Contents of the invention [0003] The object of the present invention is to provide a construction method and application of an improved adenovirus and its system, which solves the problem that the virus particles in the adenovirus system in the prior art express fiber protein and the fusion protein of fiber and target protein mixed viru...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/01C12N15/861C12N15/66A61K48/00A61P35/00
CPCA61K48/0025C12N7/00C12N15/66C12N15/86C12N2710/10021C12N2710/10043
Inventor 易俊波张宇邹永东刘立忠
Owner SHENZHEN UNIV