Taenia multiceps enolase TmENO recombinant protein with immunizing protection

A technology of Taenia polycephalum and recombinant protein, which is applied in the direction of recombinant DNA technology, medical preparations containing active ingredients, lyase, etc., can solve the problems of complex life history and insufficient immune protection against infection

CN103131719BActive Publication Date: 2014-06-04LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
Publication Date
2014-06-04

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Abstract

The invention relates to taenia multiceps protective antigen enolase TmENO recombinant protein with immunizing protection and a prepared method and application thereof. Amino acid sequence of the recombinant protein is GenBank. Amino acid sequence of ORF sequence encoding of accession number is SEQ No.5. Gene sequence of encoding gene sequence according to escherichia coli prefered codons to optimize is SEQ No.6. The recombinant protein can obviously reduce the number of cysts parasitized in sheep brains, and immunizing protection is achieved. Related experiments show that high title antibodies can be produced after animals are immunized by the recombinant protein, and the stronger immunizing protection can be created to taenia multiceps egg attack infection. The TmENO recombinant protein is a candidate target of coenurosis vaccine development and can be used for immunoprophylaxis of the coenurosis.
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Description

technical field

[0001] The invention relates to an enolase TmENO recombinant protein, an immunoprotective antigen of Taenia polycephala, and a preparation method thereof. The invention also relates to the application of the TmENO recombinant protein of the Taenia polycephalum as a vaccine for preventing and treating cerebral polycephalus. Background technique

[0002] Cerebral coenurosis (Cerebral coenurosis), commonly known as cerebral hydatid disease, also known as vertigo or staggering disease, is a tapeworm ( Taenia multiceps ) of the midtape larval brain polycephalia ( Coenurus cerebralis ) is a serious and fatal parasitic disease caused by parasitizing in the brains of ungulate ruminants such as sheep and goats. Humans can also be infected as intermediate hosts. Its adult worms live in the small intestine of canine carnivores. The lethality rate of cerebral polycephalus can reach 100%, which has caused huge economic losses to animal husbandry production. The diseas...

Examples

Embodiment Construction

[0022] 1. Preparation of recombinant protein

[0023] (1) Cloning of the TmENO gene of Taenia polycephala

[0024] 1. Genomic DNA extraction (using the tissue DNA extraction kit from Beijing Tiangen Company):

[0025] (1) Weigh 20-30 mg of the brain polycephalic protoscole, add 200 μL buffer GA, shake until the sediment is suspended.

[0026] (2) Add 20 μL proteinase K solution and mix well. Place at 56°C until the tissue dissolves, and briefly centrifuge to remove water droplets on the inner wall of the tube cap.

[0027] (3) Add 200 μL buffer GB, mix thoroughly by inversion, place at 70°C for 10 min, the solution should become clear, and briefly centrifuge to remove water droplets on the inner wall of the tube cap.

[0028] (4) Add 200 μL of absolute ethanol, vortex and mix well for 15 s. At this time, flocculent sediment may appear. Centrifuge briefly to remove water droplets on the inner wall of the tube cap.

[0029] (5) Add the solution and flocculent precipitate obt...