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Sheep coenurus skrjabini specific gene and its separation and identification

A polycephalic and specific technology, which can be used in genetic engineering, plant genetic improvement, microbial assay/inspection, etc., and can solve problems such as industrial production

Inactive Publication Date: 2007-11-14
新疆维吾尔自治区畜牧科学院兽医研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this kind of antigen cannot be produced industrially by cultivating hexacarcinoma. It can only be produced by recombinant gene vaccine. The key is to find the protective antigen gene and lay the foundation for the development of polycephalic larvae vaccine.

Method used

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  • Sheep coenurus skrjabini specific gene and its separation and identification
  • Sheep coenurus skrjabini specific gene and its separation and identification
  • Sheep coenurus skrjabini specific gene and its separation and identification

Examples

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Embodiment 1

[0049] 1. Acquisition of polycephalic stage worms of Taenia polycephalum

[0050] Collect parasitic cysts from the brain of diseased sheep, carefully separate and obtain polycephalic larvae attached to the cyst wall and suspended in the cyst fluid, wash the collected parasite sediment with sterilized PBS 10 times, and store in liquid nitrogen for later use .

[0051] 2. Extraction and RT-PCR amplification of total RNA of polycephales

[0052] Use Trizol reagent (Gibco) to extract the total RNA of polycephalus; obtain the total RNA of polycephalus, the concentration is determined to be 300ng / μl, and two clear bands of 18s and 28s can be seen by 1% formaldehyde denaturing gel electrophoresis, indicating the integrity of the total RNA good. Take 3μg RNA with SUPERSCRIPT TM A reverse transcriptase system (Invitrogen) was used to perform reverse transcription to synthesize the first strand of cDNA to obtain 20 ul of the first strand cDNA product.

[0053] The primers were desig...

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Abstract

This invention relates to one kind of sheep multi-thread you sickness specificity gene and the separation and the appraisal, the design directs the thing, with the RT-PCR method, the larva stage which the sheep multi-thread you get sick separates the new gene, its naming is 45m, its length is 698bp, codes 232 amino acids. This gene and the sheep belt tapeworm, the cow belt tapeworm's protection gene piece has the very high homology. The quantity of protein molecule expressed is approximately 6,300 road Er pause. Swells 45 milligrams to pET41b in the expression vector, the construction tallies reads the code frame's fusion gene, the expression quantity may reach 100-120mg / L . Uses in sheep's immunising experiment this protein, may arouse organism effectively the immune response, its antiserum may has the specificity response with the prosopon and the larva antigen total protein in 63KD place , It is thought that this gene has the possibility to become the candidate vaccine member which the multi-thread you get sick.

Description

technical field [0001] The present invention relates to the cloning, expression and preliminary immunity test of a new gene of polycephalic larvae in sheep, more specifically, it relates to isolating a new gene from the total RNA of polycephalic polycephalic larva The bacillus expression system expresses and purifies the immunogenic recombinant protein, which can effectively stimulate the immune response of the animal body, and its antiserum can recognize the specific protein of the natural worm (adult and larvae) antigen. Background technique [0002] Polycercariasis (commonly known as cerebral hydatid) is a zoonotic parasitic disease caused by multiceps multiceps. Adults mainly parasitize in the small intestine of the final host dog, and carnivores such as wolves and foxes are also infected. After intermediate hosts such as domestic animals, sheep, cattle, pigs, and humans eat eggs by mistake, the eggs migrate to the brain, medulla oblongata, and spinal cord, and develop ...

Claims

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Application Information

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IPC IPC(8): C12N15/12C07K14/435C12N15/63C12N1/21C12Q1/68
Inventor 由弘张文宝张壮志石保新吐尔洪·依米提王进成
Owner 新疆维吾尔自治区畜牧科学院兽医研究所
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