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Gene rEg.P29 molecular engineering vaccine resistant to sheep echinococcosis infection and preparation method and application thereof

A gene engineering vaccine, echinococcosis technology, applied in the field of gene rEg.P29 molecular engineering vaccine against sheep echinococcosis infection and its preparation

Inactive Publication Date: 2016-02-24
NINGXIA MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The technical problem to be solved by the present invention is to provide the application of a gene molecule rEg.P29 in the preparation of a genetically engineered vaccine against sheep echinococcosis infection in view of the current situation that there is no echinococcosis vaccine. Preparation

Method used

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  • Gene rEg.P29 molecular engineering vaccine resistant to sheep echinococcosis infection and preparation method and application thereof
  • Gene rEg.P29 molecular engineering vaccine resistant to sheep echinococcosis infection and preparation method and application thereof
  • Gene rEg.P29 molecular engineering vaccine resistant to sheep echinococcosis infection and preparation method and application thereof

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Experimental program
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Effect test

Embodiment Construction

[0016] Vaccine preparation:

[0017] Firstly, RNA was extracted from the Chinese strain of Echinococcus granulosus and the Eg.P29 gene was obtained by RT-PCR. The gene was included in the GenBank database with the sequence number AF078931. Secondly, the prokaryotic expression vector was constructed, and IPTG induced expression in Escherichia coli BL21. The protein rEg.P29 was identified by SDS-PAGE and Western blot, purified by His-tag affinity chromatography, and stored at -20°C.

[0018] Verification of vaccine invention effect:

[0019] 1. Preparation of experimental animals and Echinococcus granulosus eggs

[0020] Male sheep aged 4-6 months, who were negative for sheep serum (ELISA) detected by the crude antigen coating of protocercariae, were randomly divided into 3 groups: rEg.P29 vaccine group, FCA (complete Ford's adjuvant) group, PBS group, 10 rats in each group.

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Abstract

The invention discloses a gene engineering vaccine resistant to sheep echinococcosis infection. The vaccine is a protein rEg.P29 vaccine which is prepared by extracting RNA from a China strain of Echinococcus granulosus, obtaining Eg.P29 gene molecules through RT-PCR, building a P29 / pET28a prokaryotic expression vector and performing IPTG induced expression and recombination in Escherichia coli BL21. The vaccine is collected under CCTCCM2015425. Immunoprotection verification of an Echinococcus granulosus recombinant vaccine is performed on sheep, and immune protection efficiency reaches 94.8%. Vaccine candidate molecules are expected to be applied to preparing vaccines resistant to sheep echinococcosis infection and expected to play a great role in preventing and treating liver echinococcosis.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and relates to a gene rEg.P29 molecular engineering vaccine against sheep echinococcosis infection and a preparation method thereof. Background technique [0002] Echinococcus granulosus, commonly known as echinococcosis, currently has no commercially available vaccine. At present, there are more than ten kinds of candidate molecules, such as Eg95, rEg14-3-3 recombinant vaccine, ferritin recombinant vaccine, GST recombinant vaccine, etc. Among them, EG95 was first confirmed to be a sheep protective Echinococcus granulosus vaccine. The protective power of EG95 is 96-98%. Among them, domestic Kang Qiang and others have also verified the protective power of EG95 on sheep, and obtained a protective power of 90%. It is a pity that Eg95 was first successfully developed by foreign countries. Even if it is developed into a real product, its intellectual property rights do not belong to us. ...

Claims

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Application Information

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IPC IPC(8): A61K39/00A61K48/00A61P33/10C12N15/12C12N15/70
Inventor 赵巍
Owner NINGXIA MEDICAL UNIV
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