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Construction method of toxoplasma gondii SAG2 gene and ROP9 gene recombinant adenovirus, recombinant adenovirus and application

A recombinant adenovirus, gene recombination technology, applied in double-stranded DNA viruses, chemical instruments and methods, botanical equipment and methods, etc., can solve the problem of no Toxoplasma gondii vaccine and other problems

Active Publication Date: 2019-04-02
SHENYANG AGRI UNIV
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Problems solved by technology

However, there is no Toxoplasma vaccine available for humans, and there is only one veterinary Toxoplasma vaccine used in some countries: S48 attenuated strain vaccine

Method used

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  • Construction method of toxoplasma gondii SAG2 gene and ROP9 gene recombinant adenovirus, recombinant adenovirus and application
  • Construction method of toxoplasma gondii SAG2 gene and ROP9 gene recombinant adenovirus, recombinant adenovirus and application
  • Construction method of toxoplasma gondii SAG2 gene and ROP9 gene recombinant adenovirus, recombinant adenovirus and application

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Embodiment Construction

[0027] The present invention will be described in further detail below in conjunction with the accompanying drawings and specific embodiments.

[0028] Compared with other vectors, adenovirus vectors have obvious advantages. There are many reports at home and abroad on the preparation of vaccines using adenoviruses as vectors. At present, the adenovirus vector system is quite mature. The research on T.gondii is also quite clear. It has been reported that other vectors are used to develop the genetically engineered vaccine of SAG2 or ROP9, and the immune challenge test has confirmed that mice have a certain protective effect. Therefore, the present invention utilizes T.gondiiSAG2 and ROP9 The recombinant adenovirus Ad-SAG2-ROP9-EGFP was genetically constructed to develop a bivalent genetically engineered live vector vaccine for the prevention of toxoplasmosis.

[0029] The method for constructing recombinant adenovirus Ad-SAG2-ROP9-EGFP of T.gondii SAG2 gene and ROP9 gene of th...

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Abstract

The invention discloses a construction method of toxoplasma gondii SAG2 (surface antigen 2) gene and ROP9 (rhoptry protein 9) gene recombinant adenovirus, a recombinant adenovirus and an application.The construction method comprises the following steps of amplifying a Toxoplasma gondii RH strain to obtain an SAG2 gene and an ROP9 gene respectively; cloning the obtained target gene into a shuttlevector pHBAd-EF1a-MCS-3flag-CMV-EGFP; co-transfecting HEK293A cells with the recombinant adenovirus shuttle plasmid pHBAd-SAG2-ROP9-EGFP and adenovirus large skeleton pBHGlox(delta)E1, 3Cre by a liposome transfection method; and performing recombining and packaging to obtain a recombinant adenovirus Ad-SAG2-ROP9-EGFP containing the SAG2 gene and the ROP9 gene. After the recombinant adenovirus disclosed by the invention is used for immunizing Balb / c mice, the antibody level of the mice is notably increased, the proportion of activated TCL and Th lymphocytes is notably increased, cytokines of IF-gamma, IL-6, TN-alpha and the like are notably increased, and the recombinant adenovirus generates definite protective effects on the Balb / c mice.

Description

technical field [0001] The invention belongs to the field of genetic bioengineering and relates to a method for constructing a recombinant adenovirus, specifically, a method for constructing a recombinant adenovirus using Toxoplasma gondii SAG2 gene and ROP9 gene, and the recombinant adenovirus and its application. Background technique [0002] Toxoplasma gondii (Toxoplasma gondii) is an obligate intracellular nucleated parasitic protozoan. It is a serious food-borne zoonotic parasite that can infect more than 200 species of animals including livestock, poultry and humans. The human infection rate is about 25% to 50%, and there are about 500 to 1 billion people infected with toxoplasmosis in the world; the lethal outbreak of toxoplasma can cause a large number of livestock and poultry deaths, and the death rate of pigs can reach 60%. Toxoplasma gondii not only directly endangers the development of animal husbandry and product quality and safety, but also can cause public hea...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/861C12N15/12C12N15/66C12N7/01A61K39/002A61P33/02
CPCA61K39/002A61P33/02C12N7/00C12N15/86C07K14/45C12N2710/10043A61K2039/53A61K2039/70
Inventor 陈启军张东超姜宁
Owner SHENYANG AGRI UNIV
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