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Echinococcus granulosus recombinant protein EgG1Y162-2 (4) and application thereof

A technology of Echinococcus granulosus and recombinant protein, applied in the field of bioengineering, can solve the problems of non-immunogenicity, achieve good immunogenicity and specificity, and promote maturation

Pending Publication Date: 2022-08-09
XINJIANG MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the researchers believe that EgG1Y162-2 is a more ideal vaccine research target, but the problem is that the EgG1Y162-2 protein has a molecular weight of only 15kDa, and as a single small molecular protein, it is an incomplete antigen or hapten and has no immunogenicity.

Method used

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  • Echinococcus granulosus recombinant protein EgG1Y162-2 (4) and application thereof
  • Echinococcus granulosus recombinant protein EgG1Y162-2 (4) and application thereof
  • Echinococcus granulosus recombinant protein EgG1Y162-2 (4) and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Construction and identification of PET30A-EGG162-2 (4)

[0033] Specific implementation method: Permid enzyme cutting identification for the plasmid PET30A-EGG162-2 (4) synthesized by Shanghai Shengong PET30A-EGG162-2 (4) Use 1 % agar gel electrophoretic and swimming. figure 1 Essence

Embodiment 2

[0034] Example 2 Reorganization protein Egg162-2 (4) induction expression

[0035]Specific implementation method: ribbon expresses the plasmid PET30A-EGG1Y162-2 (4) Converted in ECOLI.BL21 (DE3), picked a single cloning inoculation in the LB liquid medium of 30 μg / ml of 30 μg / ml of Kenamycin. 37 ° C, 220R / min shake the bed oscillation and cultivate overnight. The next day, the medium liquid of this overnight cultivation is inached 1: 50 in the LB liquid medium containing 30 μg / ml of Kenamycin. When the mining oscillating oscillation is cultivated to 0.6-0.8, the bacteria liquid is installed in 12 50ml sterilized centrifugal tubes, and the average bacterial solution in each centrifugal tube is performed. The centrifugal tube equipped with a bacterial solution is numbered to 1 to 12, and then divided into three groups in the order of numbers. The first group 1, 2, 3, and 4 adds an inductive agent IPTG 0.2mmol / L. The second group of 5, 6, 7, and 8 adds the induction IPTG 0.5mmol / L. T...

Embodiment 3

[0036] Example 3 Nickel Pillar Purification Reorganization protein EGG1Y162-2 (4)

[0037] Specific implementation method: According to the exploring reorganized protein EGG1Y162-2 (4) The best condition for inducing, the IPTG concentration is 0.5mmol / L, a large amount of protein is induced under 37 ℃ 4 hours, collects bacteria precipitation, and per gram of bacteria per gram of bacteria Add 7ml of PBS to dilute and mix, add PMSF with a final concentration of 1mm, then perform cell ultrasonic crushing, set up working time 5s, work gap 5s, 3min throughout the process, operate three ultrasound crushing, at 4 ° C, 12000rpm / min centrifuged 10min 10min 10min 10min Get the soluble protein in the Qing Dynasty and purify the protein EGG162-2 (4) purification of the nickel column. First, use a 5x pillar volume to balance the HIS column and set aside to filter the collected protein to remove the excess impurities after removing excess impurities. The liquid over the column again. After the ...

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PUM

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Abstract

The invention discloses an echinococcus granulosus recombinant protein, which is characterized in that four EgG1Y162-2 protein fragments are connected in series through a linker sequence 'GSGGSG' to form the recombinant protein; experiments show that the recombinant protein vaccine can promote maturation of dendritic cells. The action principle and effect of the recombinant vaccine EgG1Y162-2 (4) are disclosed for the first time, and a foundation is laid for preparing vaccines or diagnostic kits for preventing and treating echinococcosis of people or livestock.

Description

Technical field [0001] The present invention is a biological engineering technology field, which involves a fine-grained echinyl tica reorganized protein EGG162-2 (4) and its application. Background technique [0002] Rhizoma disease is a severe human and animal -to -producing larvae in the middle of the middle of the dystrophilic larvae parasites that the larvae in the middle period of the naphta period in the middle of the cricket. Generally, patients are not easy to detect at the beginning of the infection. As a result, the best surgery time is missed, so most patients are mainly treated through drug treatment. Anti -parasitic drugs such as mimamazole and phenylzole are widely used in treating insect disease. Although these drugs can play a certain role, they need to be taken for a long time and have more side effects caused by the high dosage. Therefore, exploring new treatment methods is the ideal measures to prevent insecticidal diseases. A large number of studies have show...

Claims

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Application Information

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IPC IPC(8): C07K19/00C12N15/62A61K39/002A61P33/02
CPCC07K14/43554A61K39/002A61P33/02C07K2319/00Y02A50/30
Inventor 周晓涛丁剑冰赵商岐郑佳
Owner XINJIANG MEDICAL UNIV
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