Recombinant porcine parvovirus-like particle and its preparation method and application

A parvovirus, recombinant baculovirus technology, applied in biochemical equipment and methods, viruses, viral peptides, etc., can solve the problems of short duration of immunity, large immune side effects, and unsatisfactory immune effects.

Active Publication Date: 2017-10-03
GUANGXI VETERINARY RES INST
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  • Summary
  • Abstract
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  • Application Information

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Problems solved by technology

At present, inactivated vaccines developed by many units including Harbin Veterinary Research, Shanghai Academy of Agricultural Sciences, and Wuhan Keqian have been launched on the Chinese market. These vaccines have controlled the prevalence of PPV in my country to a certain extent, but they have a duration of immunity Short, low antibody level, large immune side effects and other defects, resulting in often unsatisfactory immune effect

Method used

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  • Recombinant porcine parvovirus-like particle and its preparation method and application
  • Recombinant porcine parvovirus-like particle and its preparation method and application
  • Recombinant porcine parvovirus-like particle and its preparation method and application

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Embodiment Construction

[0028] 1. Construction of fusion gene TAT-VP2

[0029] 1. Design and synthesis of primers

[0030] According to the PPV VP2 sequence published by GenBank, specific primers for amplifying the VP2 gene were designed, and the TAT protein transduction domain sequence was artificially synthesized (Table 1). The primers were synthesized by Beijing Biomed Biological Company. Among them, primers P1 and P4 are used to amplify the VP2 gene from the viral genome; P2 and P4 are used to amplify the TAT-VP2 gene introduced into the first paragraph of TAT sequence at the N-terminus of VP2, and P3 and P4 are used to amplify the TAT-VP2 gene at the N-terminus of VP2 Introduce the TAT-VP2 gene of the TAT sequence in the second paragraph, the single underlined part of P2 and P3 is the TAT sequence, and the double underlined part of P3 and P4 is the enzyme cutting site introduced.

[0031] Table 1 Target gene TAT-VP2 amplification primer and TAT protein transduction domain sequence

[0032] ...

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Abstract

The invention discloses a recombinant porcine parvovirus-like particle. A preparation method of the recombinant porcine parvovirus-like particle comprises connecting a TAT protein transduction domain and an antigen VP2 gene for forming a PPV empty capsid protein, carrying out cloning in a baculovirus transfer vector to obtain a homologous recombinant vector, carrying out packaging to obtain a recombinant baculovirus containing a PPV VP2 protein and a TAT protein transduction domain, infecting an insect cell, and expressing a recombinant TAT-VP2 protein fused with the PPV VP2 protein and the TAT protein transduction domain to obtain the parvovirus-like particle (VLPs). A research result shows that the recombinant porcine parvovirus-like particle and an adjuvant infecting an animal can effectively stimulate the PPV-specific antibody production and improve a lymphocyte production capability of lymphocyte. Therefore, the recombinant porcine parvovirus-like particle can be used to develop a novel high-efficiency PPV subunit vaccine having good immune effects.

Description

technical field [0001] The invention belongs to the technical field of porcine parvovirus, in particular to a recombinant porcine parvovirus-like particle and its preparation method and application. Background technique [0002] Porcine parvovirus (PPV) is the main pathogen that causes reproductive disorders in sows. Sows are infected by PPV in the early pregnancy, and the embryos or fetuses are invaded through the placenta, causing infertility, abortion, embryonic death, and fetal death in primiparous sows. Deformation, fetal mummification, etc., can also cause dermatitis and diarrhea in piglets, causing great economic losses to the pig industry. In recent years, PPV infection has shown an expanding and rising trend. It is prevalent throughout the country, covering more than 20 provinces and cities. The positive rate of serological surveys is generally as high as 50%-80%. PPV has become one of the important infectious diseases of pigs. First, it has greatly restricted the h...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/62C12N7/04A61K39/23A61P31/20
CPCA61K39/12A61K2039/5258A61K2039/552C07K14/005C07K2319/10C12N7/00C12N2750/14322C12N2750/14323C12N2750/14334C12N2750/14351
Inventor 吴健敏刘金凤杜毅超覃绍敏白安斌林俊陈凤莲唐海波马玲杨磊
Owner GUANGXI VETERINARY RES INST
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