Vector used for displaying antigenic epitope by using hepatitis B virus core particles, animal vaccine, and preparation method thereof

Abstract

The invention relates to a vector used for displaying antigenic epitope by using hepatitis B virus core particles, an animal vaccine, and a preparation method thereof. According to the invention, a DNA artificial synthesis method is adopted, and a coding DNA sequence of amino acid necessary for hepatitis B virus core protein is synthesized. In the synthesis process, a coding DNA sequence of hepatitis B virus core protein true peptide 1-149 site amino acid is adopted as a framework; 79-82 site amino acid coding sequence in the framework is removed, and only coding DNA relevant to virus particle assembly is preserved; for facilitating the insertion of other antigenic proteins in the hepatitis B virus core particle protein, a segment of DNA sequence coding flexible peptide is specially inserted at a position of the 79-82 site amino acid, and three endonuclease sites are introduced into the sequence, such that gene operation is facilitated; the above synthesized DNA segment is cloned into a gene cloning vector pUC18, such that a special vector pZNVC used for displaying antigenic epitope is formed; a synthesized DNA segment of an antigenic protein gene or coding epitope is inserted into pZNVC, and fusion protein is expressed in prokaryotic organisms or eukaryotic organisms. 78 amino acids on N-end of hepatitis B virus and 68 amino acids on C-end of hepatitis B virus can be folded into two columnar bodies, such that building blocks used for assembling the core protein particle are formed; flexible peptide forms a bond connecting the two columnar bodies; and proteins inserted between the flexible peptide protrude on the surface of the viral particles. The flexible peptide is composed of a series of small amino acids without side chain, and has certain flexibility. The flexible peptide does not affect the hepatitis B virus core particle protein and antigenic protein in independently and respectively folding into natural molecular structures. pZNVC is used for displaying antigenic protein antigenic epitopes, such that the existence state of the antigenic epitopes on natural pathogen outer membrane can be simulated to a maximal extent.

Description

technical field [0001] The invention relates to the field of genetic engineering DNA vaccines, in particular to a carrier for displaying antigenic epitopes by virus core particles, a vaccine and a preparation method thereof. Background technique [0002] At present, there are three main types of industrially produced vaccines: one is to kill or weaken pathogens and use them as antigens to directly manufacture vaccines; the other is to use genetic engineering technology to clone antigen genes into bacteria and produce antigenic proteins through bacterial fermentation molecules, which are then made into protein vaccines. The third type is to inject the gene encoding the antigenic protein into the body to continuously produce the antigenic protein in the body, which is DNA vaccine. However, in terms of the type of antigen, DNA vaccines are essentially protein vaccines. Among these three types of vaccines, protein vaccines are the safest, because they do not contain the geneti...

AI Technical Summary

Problems solved by technology

Second, protein vaccines are expensive to produce

However, the implementation of this technology requires the use of efficient production methods such as animal bioreactors, otherwise it is likely to be too expensive to promote

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