Dynein mosaic type recombinant human type-B adenovirus and preparation method thereof

Abstract

The invention discloses a dynein mosaic type recombinant human type-B adenovirus and a preparation method thereof. The skeleton of the dynein mosaic type recombinant human type-B adenovirus is a human type-B adenovirus genome, and a base sequence which encodes a receptor binding domain of dynein is a base sequence which encodes a corresponding domain of a human type-C adenovirus. By a molecular cloning method, Ad5-knob gene fragments are cloned and replaced to recombinant shuttle plasmids, in-vitro recombinant on the Ad5-knob gene fragments and a recombinant human type-3 adenovirus genome is realized, obtained knob gene fragments are replaced into type-5 recombinant human type-3 adenovirus genome, and therefore, dynein mosaic type recombinant human type-3 adenovirus rAd3-FK5 is obtained. The dynein mosaic type recombinant human type-3 adenovirus rAd3-FK5 can be infected with mouse primitive epithelial cells and golden hamster lung and kidney primitive cells in vitro, and the infection efficiency of the dynein mosaic type recombinant human type-3 adenovirus rAd3-FK5 is close to that of Ad5, and is much higher than that of a parent strain rAd3E, in golden hamster cells, significant copying exists, and the dynein mosaic type recombinant human type-B adenovirus can be used for small animal model research of human type-3 adenovirus vaccines and antiviral drug evaluation.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and more specifically, the invention relates to a ciliary protein chimeric recombinant human type B adenovirus and a preparation method thereof. Background technique [0002] There are more than 60 types of human adenoviruses in 7 groups. Group B adenovirus types 3, 7, 14, and 55 are the main types that cause acute respiratory infection and severe pneumonia. Adenovirus capsid mainly includes ciliary protein, penton base and hexon. It is generally believed that hexon is the main neutralizing antigen of adenovirus, and ciliary protein is the receptor-binding protein that adsorbs cells. The main domain is located at the ciliary protein knob. [0003] Appropriate animal models are required for the evaluation of adenovirus vaccines, the evaluation of the efficacy of therapeutic drugs, and the study of pathogenic mechanisms. Mice are the most commonly used model animals. However, studies ...

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Problems solved by technology

Mice are the most commonly used model animals. However, studies have shown that group B adenoviruses cannot infect mouse cells, while group C adenoviruses such as adenovirus type 5 can infect mouse cells; group B adenoviruses cannot infect golden hamster cells, Group C adenoviruses, such as type 5 adenoviruses, can infect golden hamster cells and replicate and proliferate. This difference may be caused by differences in ciliary proteins. Group B adenoviruses such as type 3 use human DSG2 protein as a receptor, while group C Adsorption receptors such as adenovirus type 5 are mainly CARs, but no studies have confirmed

At present, there is no animal model of group B adenovirus infection, so it is necessary to establish a small animal model of group B adenovirus infection

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