Tumor DNA (Deoxyribose Nucleic Acid) vaccine and virus vector vaccine taking mucoprotein 1 and surviving as targets

A DNA vaccine and anti-tumor immunity technology, applied in the field of tumor DNA vaccines and virus vector vaccines, can solve the problems of weak immunogenicity of DNA vector vaccines, relatively large safety hazards and difficult preparation of recombinant poxvirus vaccines in primary vaccination

CN104013973AInactive Publication Date: 2014-09-03CHANGCHUN BCHT BIOTECH
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
Publication Date
2014-09-03
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention relates to the field of tumor DNA (Deoxyribose Nucleic Acid) vaccines and virus vector vaccines, and in particular relates to recombinant DNA vectors VR-S8 and VR-MS, recombinant adenoviruses AD-S8 and Ad-MS and a recombinant poxvirus MVA-MS vaccine, as well as application of DNA vaccines and optimized combination of immunization ways between virus vector vaccines to preparation of antitumor vaccines.
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Description

[0001] This application is a divisional application, the original application date is December 4, 2009, the application number is 200910252427.X, and the name is "tumor DNA vaccine and virus vector vaccine targeting mucin 1 and survivin" . technical field

[0002] The invention relates to the field of tumor DNA vaccine and virus vector vaccine. Specifically, the present invention relates to recombinant DNA vector VR-S8 and VR-MS, recombinant adenovirus Ad-S8 and Ad-MS and recombinant poxvirus MVA-MS vaccine, and DNA vaccine and viral vector vaccine and viral vector vaccine The application of the optimized combination of immunization methods in the preparation of anti-tumor vaccines. Background technique

[0003] Survivin is a member of the inhibitor of apoptosis protein (IAP) family, has dual functions of anti-apoptosis and regulation of cell division, and is widely expressed in various embryonic tissues and cancer cells, but not in normal terminally differentiated cells ....

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Embodiment Construction

[0033] 1. The preparation method and conditions of the above-mentioned fragment 33M and S8

[0034] Preparation of 1.33M

[0035] MUC1VNTR is a repeat fragment containing 60 bases, GenBank number is NM_002456 (SEQ ID NO: 1). Two pairs of primers were designed according to the base sequence of a VNTR, and a VNTR (1m) repeat region fragment was synthesized by overlapping extension PCR (SOE PCR), and then digested with restriction endonucleases SalI and XhoI to produce the same sticky end characteristics, sequentially connected to construct 33M gene fragments. The specific method is as follows:

[0036] 1) Overlap extension PCR technology

[0037] The PCR reaction conditions are: 95°C for 20s, 55°C for 20s, 72°C for 30s, 30 cycles, using primers P1 (SEQ ID NO:2) and P2 (SEQ ID NO:3) to amplify one copy of MUC1VNTR without ATG (abbreviated as m), using P2 and P3 (SEQ ID NO: 4) as primers to amplify one copy of ATG-containing MUC1VNTR (abbreviated as Am) fragment.

[0038] 2) ...