Toxoplasma gondii nano-material subunit vaccine as well as preparation method and application thereof

A subunit vaccine and nanomaterial technology, which is applied in the field of Toxoplasma gondii nanomaterial subunit vaccine and its preparation, can solve the problems of vaccine spread, application limitation, and inducing serious diseases.

Pending Publication Date: 2020-06-12
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, live attenuated vaccines may spread due to virulence recovery, and may even induce severe diseases in some immunocompromised individuals, limiting their application
In recent years, studies on Toxoplasma gondii DNA vaccines

Method used

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  • Toxoplasma gondii nano-material subunit vaccine as well as preparation method and application thereof
  • Toxoplasma gondii nano-material subunit vaccine as well as preparation method and application thereof
  • Toxoplasma gondii nano-material subunit vaccine as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1 Preparation of Toxoplasma gondii nanomaterial subunit vaccine rTgPSMB1-CS

[0040] 1. Preparation of genetically engineered bacteria

[0041] Extract the total RNA of Toxoplasma gondii RH strain according to the instructions of the RNA extraction kit of Omega Company, and reverse transcribe the extracted RNA into cDNA according to the instructions of the reverse transcription kit of TaKaRa Company, using SEQ ID NO.2 and SEQ ID NO .3 primers for PCR amplification. The PCR system is as follows:

[0042]

[0043] The PCR program is as follows:

[0044]

[0045] After the PCR product was subjected to agarose gel electrophoresis at 110V for 30 minutes, the amplified 768bp band was gel-cut and recovered according to the instructions of the gel recovery kit from OMEGA Company, and the EcoR I and Hind III restriction endonucleases from TaKaRa Company were used. Instructions Carry out double enzyme digestion on the empty vector pET-32a, and refer to Novizyme’s...

Embodiment 2

[0052] Example 2 Study on the immune protection of Toxoplasma gondii nanomaterial subunit vaccine 1

[0053] 1. Experimental design

[0054] All the experimental schemes in the experimental research of this project are in compliance with the relevant regulations of Nanjing Agricultural University on animal ethics and welfare, and in line with the animal welfare protection regulations of the Science and Technology Department of Jiangsu Province.

[0055] SPF-grade male BABL / c mice weighing 18-22 g were kept in an isolation barrier system from birth to the end of the experiment in an environment free of Toxoplasma gondii, with free access to food and water. The mice were randomly divided into 15 groups, and the mice were immunized by subcutaneous multipoint injection with PBS (CONTROL group), rTgPSMB1 (PSMB1 group) and chitosan-coated rTgPSMB1 (PSMB1-CS group). 100 μL for mice (Table 1). On the 0th day, the 7th day and the 14th day, blood was collected from the mouse orbit to ...

Embodiment 3

[0065] Example 3 Study on the Immunoprotectiveness of Toxoplasma gondii Nanomaterial Subunit Vaccine 2

[0066] 3. Experimental design

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Abstract

The invention discloses a toxoplasma gondii nano-material subunit vaccine as well as a preparation method and application thereof. The vaccine is prepared by the following steps: coating toxoplasma gondii recombinant protein PSMB1 with chitosan to form nanoparticles; and after prokaryotic expression, coating the recombinant protein TgPSMB1 with nano-material chitosan to form a brand-new vaccine form. The recombinant protein TgPSMB1 is derived from toxoplasma gondii proteasome subunit 1, and the amino acid sequence of the recombinant protein TgPSMB1 is shown as SEQ ID NO.1. After the immune protection efficacy of the vaccine is evaluated, it is found that the vaccine can prolong the survival time of infected mice, it is indicated that the vaccine can provide part of immune protection, and the vaccine can be used for preventing animals from being infected with toxoplasmosis.

Description

technical field [0001] The invention relates to the technical field of biological veterinary medicines, and relates to a subunit vaccine of nano-materials of Toxoplasma gondii and its preparation method and application. Background technique [0002] Toxoplasma gondii has been discovered for more than 100 years. As an obligate intracellular parasite, it can infect almost all warm-blooded vertebrates, including mammals, birds, poultry, livestock, and even humans. , can cause serious public health problems. About 20% of people in the world are infected with Toxoplasma gondii. After infection, there are generally no clinical symptoms, but it may cause serious harm to pregnant women such as miscarriage, stillbirth or even congenital malformation of newborns. Toxoplasma gondii mainly infects pigs in livestock, and is characterized by high fever, dyspnea and reproductive disorders. Huge economic loss. At present, although pyrimethamine combined with sulfadiazine is used to contr...

Claims

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Application Information

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IPC IPC(8): A61K39/002A61P33/02C12N15/70C12N15/12C07K14/45
CPCA61K39/002A61P33/02C12N15/70C07K14/45A61K2039/552
Inventor 李祥瑞徐立新宋小凯严若峰于正青
Owner NANJING AGRICULTURAL UNIVERSITY
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