Molecular design of porcine parvovirus-like particle B cell epitope insertion site

Abstract

The invention relates to a molecular design of porcine parvovirus-like particle B cell epitope insertion site and belongs to the field of genetic engineering vaccine. Structure modeling of porcine parvovirus (PPV) capsid protein VP2 is performed by using bioinformatics software and the location of four extrusive Loop structures is determined by three-dimensional structure analysis. Firstly, on the basis of molecular simulation and literature support, we infer that loop 2,4 region can act as insertion sites of exogenous epitope gene. As is shown through experiments, after respective deletion of corresponding genes in PPV VP2 Loop2 (212aa-245aa), Loop (413aa-424aa) and then expression by adenovirus expression system, all recombinant virus with deletion mutations of Loop 2,4 can assemble regular virus-like particles [PPV: delta V LPs]. The invention also relates to an application of recombinant PPV delta V P2 virus-like particles of exogenous gene expressed by the recombinant virus in vaccination and the like.

Description

1. Technical field [0001] The invention relates to the molecular design of the porcine parvovirus-like particle B cell epitope insertion site, and belongs to the field of genetic engineering vaccines. 2. Background technology [0002] The porcine parvovirus (PPV) virion is hexagonal or circular in appearance, has no envelope, and isometrically symmetrical to an icosahedron. The capsid is composed of 32 capsomers, and the PPV genome encodes 2 structural polypeptides, VP1 and VP2. The molecular masses are 84ku and 64ku, respectively, and another structural polypeptide VP3 is produced by the hydrolysis of VP2, with a molecular mass of 60ku. Among them, VP2 is the main capsid protein constituting the virion, which contains the main protective antigen of PPV. The C-terminal amino acid sequence of VP1 protein overlaps with the N-terminal amino acid sequence of VP2. The C-terminus of VP2 is exposed on the surface of the protein, so the integrity of the C-segment is necessary to m...

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Problems solved by technology

According to this analysis, it is believed that although the N-terminal of VP2 can be used as the insertion site to form virus-like particles, but the N-terminal of the empty particles is in the cylindrical channel of the quintuple axis, and the recombinant cannot stimulate the immune response to produce B cell antigen epitopes; Peptides that form canyons around the quintuple axis and depressions in the direction of the biad axis are also not suitable as insertion sites

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