Replication-defective A virus expression vector system and vaccine preparation method

A technology for expressing vectors and alphaviruses, which is applied in the direction of antiviral agents, viral antigen components, and the use of vectors to introduce foreign genetic materials, etc. It can solve the problems of lack of vaccine preparation methods, and achieve the effect of avoiding inability to expand cultivation and reducing the risk of use

Inactive Publication Date: 2012-05-09
SOUTH CHINA UNITED VACCINE INST
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  • Replication-defective A virus expression vector system and vaccine preparation method
  • Replication-defective A virus expression vector system and vaccine preparation method
  • Replication-defective A virus expression vector system and vaccine preparation method

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preparation example Construction

[0034] A vaccine preparation method, comprising the following steps:

[0035] 1) Insert the immunogen sequence of the germ into the replicon of the alphavirus replicon system described in claim 1 or 3, replace the structural protein sequence of the original alphavirus nucleic acid sequence, and obtain the immunogen replicon;

[0036] 2) The structural protein helper of the alphavirus auxiliary packaging system described in claim 1 or 3 is transfected into the cell expressing the gene replication regulatory switch nucleic acid sequence inhibitory protein to obtain the structural protein auxiliary cell;

[0037] 3) Adding a substrate that binds to the nucleic acid sequence inhibitory protein of the gene replication regulatory switch in the culture medium of the structural protein auxiliary cell to relieve the inhibition of the cell on the expression of the structural protein;

[0038]4) Use the immunogen replicon to transfect the helper cells that relieve the inhibition of struc...

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Abstract

The invention discloses a replication-defective A virus expression vector system and a vaccine preparation method. The vector system consists of a replication-defective A virus vector of a deleted structural gene and minus strand complementary plasmids or cells containing a controllable virus A structural gene. In the preparation method, an exogenous gene target is introduced by constructing the replication-defective A virus vector of a deleted virus structural protein; and the replication-recombinant A virus can be continuously produced in a large scale by combining the minus strand complementary plasmids or cells containing the controllable A virus structural gene, so that the problem of low virus yield of a conventional A virus system can be solved, a target of preparing recombinant virus in a large scale is realized, and exogenous protein and virus sample particles can be expressed in scale. The replication-defective A virus expression vector system disclosed by the invention is suitable for large-scale production of DNA (deoxyribonucleic acid) vaccine, vector virus vaccine and exogenous protein expression.

Description

technical field [0001] The invention relates to an alphavirus expression vector system and a method for preparing a vaccine using the expression system, in particular to a method for large-scale cultivation of a recombinant alphavirus of a replication-deficient alphavirus expression vector system, and a method for preparing a vaccine using the expression system. Background technique [0002] The nucleic acid sequences of alphaviruses in the Togaviridae family are mainly composed of two parts, structural and nonstructural protein sequences. Replicons with non-structural protein sequences can replicate RNA, and can amplify the subgenome of structural genes in large quantities, and can express a large amount of exogenous proteins that replace structural genes. However, due to the absence of structural genes, it cannot be individually packaged into infectious virus particles, and cannot reinfect cells. Taking advantage of this feature, replication-defective alphaviruses can be ...

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Application Information

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IPC IPC(8): C12N15/86A61K39/00A61K39/125A61K39/13A61P31/14
CPCY02A50/30
Inventor 彭涛刘钿莲王弋
Owner SOUTH CHINA UNITED VACCINE INST
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