Recombinant adenovirus high-expression vector for promoting effective presentation of Hantavirus fusion protein G2S0.7

Abstract

The invention relates to a recombinant adenovirus high-expression vector for promoting effective presentation of Hantavirus fusion protein G2S0.7. A G2S0.7-pCAG transfer vector containing G2S0.7 mosaic gene of Hantavirus 76-118 strain and a CAG promoter / enhancer is mainly transformed to promote the effective presentation of the high-expression Hantavirus fusion protein Hantavirus in an organism. By a gene recombination technology, the recombinant adenovirus transfer vector G2S0.7-pCAG containing the mosaic gene G2S0.7 is transformed, and Ub gene is connected to the vector. The recombinant transfer vector and the adenovirus vector are respectively subjected to double digestion, and a recombinant fragment Ub-G2S0.7-pCAG is integrated into the DNA of the adenovirus vector; and after packaging, purification and titer detection, rAd-Ub-G2S0.7-pCAG and recombinant adenovirus rAd-G2S0.7-pCAG containing the G2S0.7 mosaic gene and the CAG promoter / enhancer, which is constructed early by the inventor are respectively used for immunizing C57BL / 6 mice, immunological characteristics are researched, and results show that compared with the conventional recombinant adenovirus, the recombinant adenovirus can effectively improve partial humoral immune response level and partial cellular immune response level of organisms of experimental animals.

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 G2S0.7 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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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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