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Recombinant adenovirus high expression vector pCAG-G1S0.7-HSP70C for promoting effective presentation of hantavirus fusion protein G1S0.7

A recombinant adenovirus, fusion protein technology, applied in the introduction of foreign genetic material using a vector, recombinant DNA technology, applications, etc., can solve the problems of low protein expression and unsatisfactory immune level.

Inactive Publication Date: 2014-08-13
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

Method used

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  • Recombinant adenovirus high expression vector pCAG-G1S0.7-HSP70C for promoting effective presentation of hantavirus fusion protein G1S0.7
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  • Recombinant adenovirus high expression vector pCAG-G1S0.7-HSP70C 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 (Catalog 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 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 ligated with the T-easy carrier overnight at 14°C, and the...

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Abstract

The invention relates to a recombinant adenovirus high expression vector pCAG-G1S0.7-HSP70C for promoting effective presentation of hantavirus fusion protein G1S0.7 containing heat shock protein HSP70C, which is mainly used for modifying a G1S0.7-pCAG transfer vector of G1S0.7 chimeric genes and CAG starters / enhancers containing 76 to 118 strains of hantaviruses and is used for promoting the effective presentation of the high-expressed hantavirus fusion protein G1S0.7. A genetic recombination technology is used for modifying the recombinant adenovirus transfer vector G1S0.7-pCAG containing the chimeric gene G1S0.7, and the end of an HSP70 antigen presentation molecular gene C is connected to the vector. Due to the adoption of the recombinant adenovirus high expression vector, the presentation efficiency of the fusion protein G1S0.7 in an animal body can be effectively improved, and the body humoral immunity response level and the cell immunity response level can be 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 the recombinant adenovirus high-expression vector pCAG- G1S0.7-HSP70C. 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 high mortality rate. So far, there is still a lack of specific and effective therapeutic drugs in clinical pract...

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Application Information

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Patent Type & Authority Patents(China)
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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