Recombinant multivalent vaccine

A virus genome and varicella zoster technology, applied in the field of recombinant varicella zoster virus, can solve the problem of inserting multiple antigens into the virus genome

Inactive Publication Date: 2009-02-04
THE RES FOUND FOR MICROBIAL DISEASES OFOSAKA UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Also, when using BAC vectors to prepare multivalent vaccines, there is also the problem that multiple antigens must be inserted into the viral genome

Method used

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  • Recombinant multivalent vaccine
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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0330] (Example 1: Selection of a region (gene) to be inserted into a BAC vector)

[0331] When using BAC vectors to prepare recombinant viruses inserted with foreign antigen genes, the genome size increases due to the insertion of many foreign genes. It is known that if the genome size is too large, the genomic DNA cannot be packaged into the capsid, and as a result, recombinant virus cannot be prepared. Therefore, in order to insert most of the antigen genes into the Oka vaccine strain, knockout of non-essential genes of the Oka vaccine strain must be considered to reduce the size of the genomic DNA. In addition, a non-essential gene that can grow even if the virus is knocked out is suitable as a site for inserting a foreign sequence such as a BAC vector sequence or a gene encoding an antigenic protein derived from another virus.

[0332] Therefore, using the recombinant DNA (P-Oka strain VZV-BAC-DNA) obtained by inserting the genome of the original strain of varicella viru...

Embodiment 2

[0339] (Example 2: Preparation of a multivalent vaccine by inserting the mumps virus HN gene into the ORF of gene 13)

[0340] As shown in Example 1, gene 13 is a suitable gene for knockout and / or insertion of foreign sequences, therefore, a multivalent vaccine is prepared by inserting the mumps virus HN gene into the ORF of gene 13.

[0341] The HN gene and F gene of mumps virus were amplified from the wild strain-Iwasaki strain using PCR. Analyzing the amino acid sequences of the F gene and HN gene of the cloned Iwasaki strain, the F gene showed higher homology (>98.5) compared with the field strain or the vaccine strain, while the HN gene had a higher homology (>98.5) with the field strain after the late 1990s. The homology of the strains is high, and the homology with field strains or vaccine strains before the early 1990s is low (about 96%).

[0342] Next, a human cytomegalovirus (CMV) promoter / enhancer sequence was operably linked upstream of the cloned gene. The plasm...

Embodiment 3

[0349] (Preparation of mutant recombinant varicella-zoster virus strain with weak pathogenicity)

[0350] According to the present invention, a mutant recombinant varicella-zoster virus can be prepared by adopting the following method, and a mutant varicella-zoster virus strain with weak pathogenicity can be obtained from the mutant virus.

[0351] (1: Preparation of mutant recombinant varicella-zoster virus)

[0352] The preparation method of the mutated recombinant varicella-zoster virus includes, for example: preparing the mutated recombinant varicella-zoster virus by homologous recombination between the nucleic acid containing the mutated gene and the VZV-BAC-DNA plasmid. The mutated gene used in homologous recombination with the VZV-BAC-DNA plasmid may have random variation or directed site variation. Using any one of the above methods, a mutant recombinant varicella-zoster virus group with random variation and a mutant recombinant varicella-zoster virus with directed si...

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PUM

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Abstract

Disclosed are recombinant varicella-zoster virus, a process for producing the virus, a pharmaceutical composition comprising the recombinant varicella-zoster virus, a vector having a BAC vector sequence in a specific gene of the varicella-zoster virus genomic gene, a cell having the vector therein, a fragment capable of homologous recombination with the varicella-zoster virus genome, a nucleic acid cassette comprising a BAC vector sequence, and a polyvalent vaccine. Provided is a process for producing a recombinant varicella-zoster virus having a BAC vector sequence inserted in a specific viral gene.

Description

technical field [0001] The present invention relates to a recombinant varicella-zoster virus, especially a recombinant varicella-zoster virus prepared by using BAC (Escherichia coli artificial chromosome), and a pharmaceutical composition containing the virus. The present invention further relates to a vector containing varicella-zoster virus genome gene and BAC vector sequence, and a cell containing the above-mentioned vector. The present invention also relates to a method for preparing recombinant varicella-zoster virus. The present invention also relates to a nucleic acid cassette comprising a segment capable of homologous recombination with the varicella-zoster virus genome, and a BAC vector sequence. Background technique [0002] Varicella-zoster virus (VZV), a virus belonging to the family Human Herpesviridae, is the cause of diseases (chickenpox and herpes zoster) that exhibit two distinct clinical manifestations. Initial infection with the virus causes chickenpox. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/00A61K39/25A61P31/22A61P37/04C12N5/00C12N7/00
CPCA61K39/165A61K2039/70C12N2800/204A61K39/25C12N2710/16734A61K39/12C12N15/86C12N2820/55C12N2760/18734C12N2710/16762C12N2800/50C12N2710/16743A61K39/295A61K2039/5256A61K2039/5254A61P31/04A61P31/12A61P31/22A61P37/04Y02A50/30
Inventor 森康子P·索姆布恩图姆吉井洋纪五味康行高桥理明山西弘一
Owner THE RES FOUND FOR MICROBIAL DISEASES OFOSAKA UNIV
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