AIDS vaccine and preparation method thereof

A technology of AIDS and vaccines, applied in the field of genetic engineering, can solve the problems of high cost, high price, and limited effect, and achieve the effects of saving time, easy cultivation, and rapid growth

Inactive Publication Date: 2018-11-16
GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, HAART treatment is expensive and less feasible for African countries with more severe HIV epidemics and developing countries [1] ; Moreover, HAART therapy can only control the replication of the virus, but cannot completely remove the virus. In addition, it is expensive, has limited effects and has serious side effects

Method used

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  • AIDS vaccine and preparation method thereof
  • AIDS vaccine and preparation method thereof
  • AIDS vaccine and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] The construction of embodiment 1 recombinant plasmid

[0041] (1) Construction of integrated plasmid pI1818gp, plasmid pI2224gp, plasmid pI3763gp and intracellular control plasmid pIgp, the structure diagrams are shown in figure 1 , figure 2 , image 3 and Figure 4 , see the construction process respectively Figure 4 and Figure 5 (The construction flowchart of plasmid pI2224gp and plasmid pI3763gp can be deduced by analogy). Depend on figure 1 As shown, plasmid pI1818gp contains resistance selection gene AmpR, Escherichia coli replicon oriE, promoter hsp60, carrier protein gene Rv1818c, Linker sequence, antigen gene gp120 derived from simian immunodeficiency virus SIV, and human influenza blood in a clockwise direction. The lectin virus gene tag HA-tag, the fusion gene of the resistance screening gene Hyg and the integrase gene Int, and the fusion gene of the phage integration site Attp and the integrase gene Int; figure 2 As shown, the plasmid pI2224gp has ...

Embodiment 2

[0071] The construction of embodiment 2 recombinant Mycobacterium smegmatis

[0072] The relevant plasmid constructed in Example 1 was transformed into Mycobacterium smegmatis ZWY2, and the expression of the antigenic protein gene gp120 in each strain was verified by western-blot, and it was expressed on the surface of Mycobacterium smegmatis ZWY2 strain. The specific operation is as follows:

[0073] 1. Conversion:

[0074] The plasmids pI1818gp, pI2224gp, pI3763gp and pIgp were transformed into Mycobacterium smegmatis ZWY2 by electroporation (see reference 18 for the transformation method), and corresponding transformants were obtained, which were respectively named as pI1818gp strain, pI2224gp strain, The pI3763gp strain and the pIgp strain, wherein, the pI1818gp strain, the pI2224gp strain, and the pI3763gp strain are AIDS vaccines.

[0075] 2. Western-blot verification of the surface display of the antigenic protein gene gp120:

[0076] Firstly, the subcellular compone...

Embodiment 3

[0081] Embodiment 3 AIDS vaccine immunogenicity test

[0082] Evaluate the immunogenicity of embodiment 2 AIDS vaccine by animal experiments, 84 6-week-old C57BL / 6 female mice purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd. were randomly divided into 7 groups, respectively pI1818gp strain group, pI2224gp strain group, pI3763gp strain group, pIgp strain group, ZWY2 group, PBS group (negative control) and adenovirus vector Ad5SIV-env group (positive control, gifted by researcher Chen Ling, Guangzhou Institute of Biomedicine and Health, Chinese Academy of Sciences), each The number of animals in the group is shown in Table 1. The IACUC number of this animal experiment is 2017024.

[0083] Table 1 Mouse grouping and immunization rules

[0084]

[0085] The specific operation is as follows:

[0086] 1) Cell preparation and immunization, the specific operation is as follows:

[0087] a. Inoculate the bacterial vaccines pI1818gp strain, pI2224gp st...

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Abstract

The invention discloses a recombinant plasmid. The recombinant plasmid contains a gp120 gene of the monkey immunodeficiency virus and at least one cell wall carrier protein gene of Mycobacterium tuberculosis. The invention also discloses recombinant mycobacterium smegmatis which can stably express the gp120 gene of the monkey immunodeficiency virus and at least one cell wall carrier protein gene of Mycobacterium tuberculosis. The invention also discloses an AIDS vaccine and a preparation method thereof. The AIDS vaccine contains the recombinant mycobacterium smegmatis, is easy to culture, grows fast and saves time, labor, material resources and financial resources. The vaccine comprising three strains of the recombinant mycobacterium smegmatis ZWY2 exhibiting the SIV gp120 antigen proteinon the surface can stably exhibit the SIV gp120 antigen protein on the surface of the mycobacterium smegmatis ZWY2 and has the immunological effect in C57 mice far superior to that of intracellular expression of the antigen protein gp120 and that of the already published AIDS vaccine exhibiting gp120 on the surface based on adenovirus vector construction.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, and also relates to an AIDS vaccine, in particular to a recombinant mycobacterium smegmatis obtained through biotechnological transformation. Background technique [0002] Since the US Centers for Disease Control and Prevention reported the world's first AIDS case in 1981, researchers have been working on developing a safe and effective HIV vaccine to prevent and treat AIDS. At present, the most famous and effective HIV treatment is cocktail therapy, namely highly active antiretroviral therapy (Highly Active Anti-Retroviral Therapy, HAART), which is a combination of three or more antiviral drugs for treatment. AIDS. However, HAART treatment is expensive and less feasible for African countries with more severe HIV epidemics and developing countries [1] ; And, HAART therapy can only control the virus replication, can not completely remove the virus, in addition to the high price, limi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/74C12N15/66C12N15/49C12N15/31C12N1/21A61K39/21A61P31/18C12R1/34
CPCA61K39/12A61P31/18C12N15/66C12N15/74C07K14/005C07K14/35A61K2039/523C12N2740/15034C12N2740/15022
Inventor 张天宇刘燕孙彩军潘恩祥王邦兴郭灵敏陈凌刘志勇
Owner GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI
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