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Recombinant adenovirus high expression vector for promoting effective presentation of hantavirus fusion protein G1S0.7

A technology of recombinant adenovirus and fusion protein, which is used in the fields of recombinant adenovirus high expression vector, immunology and immune applications, and molecular biology, and can solve the problems of unsatisfactory immune level and low protein expression.

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. For example, 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

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  • Recombinant adenovirus high expression vector for promoting effective presentation of hantavirus fusion protein G1S0.7
  • Recombinant adenovirus high expression vector for promoting effective presentation of hantavirus fusion protein G1S0.7
  • Recombinant adenovirus high expression vector for promoting effective presentation of hantavirus fusion protein G1S0.7

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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 G1S0.7-pCAG used in the present invention was previously constructed by the applicant. (see patent application number 200910254561.3)

[0027] The recombinant adenovirus rAd-G1S0.7-pCAG and rAd-G1S0.7 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 Ub gene sequence inserted in the present invention is Ub up primer, Ub down primer upstream and downstream primers, pCMV-F3Ub as template, and 55°C as annealing temperature for PCR amplification, as shown in the attached figure 1 shown. The PCR product was ligated with the T-easy carrier overnight at 14°C, and the ligat...

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Abstract

The invention relates to a ubiquitin Ub-containing recombinant adenovirus high expression vector for promoting effective presentation of a hantavirus fusion protein G1S0.7. Primarily, G1S0.7 mosaic genes of hantavirus-containing 76-118 strains and a G1S0.7-pCAG transfer vector of a CAG (Cytotoxin Associated Gene) promoter / enhancer are constructed for promoting effective presentation of highly expressed hantavirus fusion protein G1S0.7 by an organism. Compared with the recombinant adenovirus, partial humoral immunity response level and partial cell immunity response level of an animal organism 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 invention relates to the reconstruction of an adenovirus recombinant transfer vector and a recombinant adenovirus high-expression vector for promoting the effective presentation of hantavirus fusion protein G1S0.7 to the pathogen of hemorrhagic fever with renal syndrome (HFRS). 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 of patients, and a...

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Application Information

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IPC IPC(8): C12N15/861C12N15/66
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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