Preparation method and application of taenia pisiformis 18kDa antigen

A tapeworm and bean-like technology is applied in the field of preparation of 18 kDa antigen of Taenia bean, which can solve the problems of limited sources of natural worm antigens and difficulty in popularization and use.

Inactive Publication Date: 2013-05-01
SICHUAN AGRI UNIV
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Problems solved by technology

[0003] The present invention provides a preparation method and application of the 18 kDa antigen of Taenia fabaciformis, aiming to solve the problem that the natural insect antigen is affected by the source of the tapeworm when screening the protective antigen of the insect body itself to prepare a vaccine to block the transmission route of the tapeworm. Restrictions, problems that are difficult to promote and use clinically

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preparation example Construction

[0056] The preparation method comprises the following steps:

[0057] Step S101, activating Taenia lentilis, and extracting total RNA;

[0058] Step S102, using total RNA as a template to synthesize cDNA;

[0059] Step S103, amplifying the Tp18 coding region, and performing bioinformatics analysis of the Tp18 gene;

[0060] Step S104, performing prokaryotic expression of Tp18, and purifying the expressed protein.

[0061] In the embodiment of the present invention, in step S101, the implementation method of activating Taenia fauna and extracting total RNA is as follows:

[0062] Collect fresh Cysticercus lentiformis-infected dogs from naturally infected rabbits, collect proglottis five weeks later, wash them with PBS buffer, and store them in a preservation solution at 4°C;

[0063] The collected 200 segments were washed several times with PBS buffer, cut into pieces in a 1.5 mL centrifuge tube, incubated with 1% pepsin solution at 37°C for 1 h, and centrifuged at 3000 r / mi...

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Abstract

The invention belongs to the technical field of biology of genes, and provides a preparation method and an application of a taenia pisiformis 18k Da antigen. Taenia pisiformis hexacanth embryos are activated and subjected to total RNA (Ribose Nucleic Acid) extraction; the total RNA is taken as a template to synthetize a cDNA; a Tp18 code area is amplified, and the Tp18 gene bioinformatics analysis is carried out; the prokaryotic expression is performed on the Tp18, and the expression protein is purified. According to the preparation method, the Tp18 genes are amplified from the activated aenia pisiformis hexacanth embryos by an RACE (rapid-amplification of cDNA ends) PCR (Polymerase Chain Reaction) technology to construct a pET32a / Tp18 expression vector, the recombination protein is obtained through IPTG induction, and the obtained recombination protein has better immunogenicity, can serve as a candidate antigen of a taenia pisiformis vaccine, and has stronger popularization and application values.

Description

technical field [0001] The invention belongs to the technical field of gene biology, and in particular relates to a preparation method and application of the 18 kDa antigen of Taenia lentils. Background technique [0002] Taenia lentilis is a common parasite of canids (terminal host) and lagomorpha (intermediate host), among which cysticercosis lentilis caused by tapeworm stage larvae has brought huge economic losses to the rabbit breeding industry . Taenia lentils is distributed worldwide. Adults parasitize the small intestine of final hosts such as dogs and foxes. The middle tapered larva, Cysticercus lentils, parasitizes the liver capsule and gastric reticulum of lagomorpha animals such as rabbits. Parts such as membrane and mesentery can cause rabbits to become thin and disease resistant, and even die. China is a big country in raising rabbits in the world. Cysticercosis beaniformis is widely prevalent in our country, which brings huge economic losses to the rabbit ind...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C12N15/70C07K14/435
Inventor 杨光友杨德英彭雪蓉古小彬
Owner SICHUAN AGRI UNIV
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