Recombined prokaryotic expression detection antigen for detecting local trichina nematode antibody and producing method thereof

A Trichinella elegans, prokaryotic expression technology, applied in biochemical equipment and methods, measuring devices, biological testing, etc., can solve problems such as low detection rate, difficulty in animal quarantine work, and patient pain

Inactive Publication Date: 2006-08-02
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Abstract
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  • Claims
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Problems solved by technology

[0003] Because the trichinella worms are very small, for a long time, the diagnosis of trichinosis has been mainly based on microscopic examination of the diaphragm after slaughter or using artificial pepsin to digest the muscle and collect the worms for detection. Although it is relatively intuitive, it is very cumbersome and the detection rate Low, and can not be diagnose

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Embodiment Construction

[0033] The present invention is a gene recombinant prokaryotic expression protein of a main protective antigen 49ku excreted and secreted by local Trichinella nematodes during the growth process. The TNPG gene contains 951 bases and encodes 316 amino acid residues. When expressed in the prokaryotic, the TNPG gene and The expression vector pET-30a was fused and expressed, and the molecular weight of the fusion protein was 40.8KU.

[0034] The amino acid sequence of the prokaryotic expression protein of the local Trichomona TNPG gene recombinant prokaryotic expression protein of the product of the present invention is (316 amino acid residues in total):

[0035] Product preparation process of the present invention:

[0036] (1) Design and amplify the specific primers of the local Trichomus TNPG gene;

[0037] (2) obtain the TNPG gene fragment by PCR amplification;

[0038] (3) Cloning and sequencing identification of the TNPG gene;

[0039] (4) directional subcloning of TNPG ...

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Abstract

The present invention provides a recombinant prokaryotic expression protein antigen for detecting autochthonous trichinella spiralis antibody and its preparation method. Said product can be used as detection antigen of indirect-ELISA and Dot-ELISA method for detecting autochthonous trichinella spiralis antibody, also can be used as gene engineering subunit vaccine for preventing autochthonous trichinella spiralis infection. The preparation process of said product includes the following steps: (1) designing specific primer for amplifying TNPG gene; (2), utilizing PCR amplification to obtain TNPG gene segment; (3), cloning TNPG gene and sequencing, and making identification; (4), directionally subcloning TNPG gene to pET-3a prokaryotic expression vector; (5), induced expression of TNPG gene in colibacillus; and (6), affinity chromatography purification of TNPG gene expression protein.

Description

technical field [0001] The invention relates to a preparation method of a genetic engineering product, in particular to a TNPG gene recombination prokaryotic expression detection antigen protein for detecting native trichinella trichinella antibodies and its preparation method. Background technique [0002] Trichinellosis (Trichinellosis) is an important zoonotic nematode disease caused by the adult and larvae of Trichinella spiralis infecting humans or animals. It occurs all over the world, and the local Trichinella nativa is one of its pathogens . The disease is mainly caused by eating raw or undercooked pork or other animal meat containing Trichinella cysts, and causes human death due to myocarditis and pneumonia. Because trichinella has many infected hosts, the prevalence of trichinellosis is wide and the transmission route is complicated, so this serious threat to human health is caused by animal husbandry (especially pig farming), meat industry and foreign trade expor...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N33/531C12Q1/00
Inventor 宋铭忻郑宝亮路义鑫李冬梅韩周马广鹏武嘉男
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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