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Recombinant adenovirus expression vector containing heat shock protein HSP70C for promoting hantavirus fusion protein G2S0.7 presentation

A technology of recombinant adenovirus and heat shock protein, applied in application, biochemical equipment and methods, botany equipment and methods, etc., can solve the problems of low protein expression and unsatisfactory immune level

Inactive Publication Date: 2012-09-19
FOURTH MILITARY MEDICAL UNIVERSITY
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Problems solved by technology

[0005] However, we also found some problems in the research, such as although the complete fusion protein can be expressed by using various systems, the overall protein expression is still low
In addition, after immunizing animals with fusion proteins, although each expression system can stimulate the body to produce humoral and cellular immune responses, and the effect of the adenovirus expression system is better than other systems, the overall immune level is not very satisfactory

Method used

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  • Recombinant adenovirus expression vector containing heat shock protein HSP70C for promoting hantavirus fusion protein G2S0.7 presentation
  • Recombinant adenovirus expression vector containing heat shock protein HSP70C for promoting hantavirus fusion protein G2S0.7 presentation
  • Recombinant adenovirus expression vector containing heat shock protein HSP70C for promoting hantavirus fusion protein G2S0.7 presentation

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Embodiment Construction

[0025] The adenovirus expression system used in the present invention was purchased from Adeno-X of CLONTECH company TM system (Product No. K1650-1), the transfer vector is pShuttle.

[0026] The recombinant adenovirus transfer vector G2S0.7-pCAG used in the present invention was previously constructed by the applicant (see original patent application number 200910254563.2).

[0027] The recombinant adenovirus rAd-G2S0.7-pCAG and rAd-G2S0.7 used in the present invention were packaged and purified by the applicant in the early stage.

[0028] The bivalent HFRS inactivated vaccine used in the present invention was purchased from Zhejiang Tianyuan Company.

[0029] The HSP70C gene sequence inserted in the present invention uses HSP70C up primer and HSP70C down primer as primers, and uses the Mycobacterium tuberculosis genome as a template to perform PCR amplification, as shown in the attached figure 1 shown. The PCR product was connected to the T-easy vector overnight at 14°C, a...

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Abstract

The invention relates to a recombinant adenovirus expression vector containing heat shock protein HSP70C for promoting hantavirus fusion protein G2S0.7 presentation. A G2S0.7-pCAG transfer vector containing CAG promoter / enhancer and G2S0.7mosaic gene of hantavirus 76-118 strain is mainly built to promote organism to effectively present highly expressed hantavirus fusion protein G2S0.7. According to the invention, humoral immune response level and cellular immune response level of an experimental animal body can be effectively improved.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to related fields such as molecular biology, immunology and immune application. The present invention relates to the reconstruction of adenovirus recombinant transfer vector and a recombinant adenovirus high-expression vector G2S0 for promoting the effective presentation of Hantavirus fusion protein G2S0.7 to the pathogen of hemorrhagic fever with renal syndrome (HFRS). 7-pCAG. Background technique [0002] Research status of hemorrhagic fever with renal syndrome and its genetic engineering vaccine: [0003] Hemorrhagic fever with renal syndrome (HFRS) is caused by Hantavirus (HV) and is an acute viral infectious disease transmitted by rodents. Clinically, it is characterized by fever, bleeding and acute renal failure. Damage is the main feature. China is the country with the most severe HFRS epidemic in the world. It has the characteristics of a wide range of epidemics, a large number...

Claims

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Application Information

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IPC IPC(8): C12N15/861
Inventor 徐志凯张芳琳白文涛李璞媛程林峰李凯吴兴安于澜张亮
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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