Tubercle bacillus fusion protein and preparation method and application thereof

A fusion protein, Mycobacterium tuberculosis technology, applied in fusion polypeptides, chemical instruments and methods, bacteria, etc., can solve problems such as unconsidered

Inactive Publication Date: 2011-09-14
LANZHOU UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the past, fusion proteins with various tags (such as His-Tag, GST-Tag, S-Tag, etc.) were often constructed for the convenience of later purification methods, but did not consider whether the tags would affect animal experiments and Approval for further clinical trials

Method used

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  • Tubercle bacillus fusion protein and preparation method and application thereof
  • Tubercle bacillus fusion protein and preparation method and application thereof
  • Tubercle bacillus fusion protein and preparation method and application thereof

Examples

Experimental program
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Embodiment 1

[0058] 1. Construction, expression and purification of fusion protein EAMM

[0059] Fusion protein ESAT6-Ag85B-Mpt64 190-198 -Mtb8.4 is to connect and clone ESAT-6, Ag85B, 190-198 amino acid sequences of Mpt64 and Mtb8.4 gene to construct an Escherichia coli expression vector, express it in Escherichia coli, and purify the target protein.

[0060] 1. Fusion protein ESAT6-Ag85B-Mpt64 190-198 -Build of Mtb8.4(EAMM):

[0061] 1) PCR amplification of ESAT-6 gene: due to fusion expression of Ag85B-Mpt64 behind ESAT-6 gene 190-198 -Mtb8.4, so the ESAT-6 termination signal was removed when designing the primers, and the ESAT-6 fragment was amplified using the 5'-end specific primer ESAT-6F and the 3'-end primer ESAT-6R. The PCR reaction conditions were as follows: pre-denaturation at 96°C for 1 minute; denaturation at 98°C for 10 seconds, renaturation at 60°C for 15 seconds, extension at 72°C for 30 seconds, 30 cycles. After the PCR product was purified, it was digested with Nde ...

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Abstract

The invention discloses a tubercle bacillus fusion protein which is a protein with a sequence 2 in a sequence table, and provides a preparation method of the protein and application of the protein in preparation of vaccine. The constructed, expressed, purified and label removed fusion protein ESAT6-Ag85B-Mpt64190-198-Mtb8.4 (EAMM) provided by the invention aims to establish immune response aiming at tubercle bacillus to improve the protection effect of the vaccine. The constructed fusion protein EAMM and an adjuvant are mixed to form tubercle subunit vaccine, and the tubercle subunit vaccine for inducing immune response of mainly immunizing Th1 type cells and humoral immune response has strong immunogenicity and good anti-tubercle protection effect.

Description

technical field [0001] The invention relates to a mycobacterium tuberculosis fusion protein and its preparation method, as well as its application in the preparation of tuberculosis subunit vaccine. Background technique [0002] Mycobacterium tuberculosis is the pathogenic bacterium that causes tuberculosis in humans and animals. Mycobacterium tuberculosis faces great difficulties in drug treatment due to its unique biological characteristics, such as thick cell wall, long growth cycle, latent infection and easy drug resistance. Although in the first half of the last century, Bacillus Calmette-Guerin (BCG) for the prevention of tuberculosis and specific drugs for the treatment of tuberculosis (such as isoniazid, rifampicin, etc.) were successfully developed, but by the end of the last century, tuberculosis was still a global infection and infection. The number one "killer" of the disease. Moreover, with the ravages of AIDS (HIV) in the world and the emergence of multi-drug ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/63C12N1/21A61K39/04A61P31/06
CPCC07K14/35A61K39/04C07K2319/00A61P31/06
Inventor 祝秉东辛奇胡丽娜王秉翔达泽蛟牛红霞刘万波唐克峰高娃景涛
Owner LANZHOU UNIVERSITY
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