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Tubercle bacillus recombined protein and expressing purefying method and application thereof

A technology of recombinant protein, Mycobacterium tuberculosis, applied in chemical instruments and methods, biochemical equipment and methods, applications, etc.

Inactive Publication Date: 2006-06-28
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] 2. Live attenuated vaccine for auxotrophic tuberculosis,

Method used

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  • Tubercle bacillus recombined protein and expressing purefying method and application thereof
  • Tubercle bacillus recombined protein and expressing purefying method and application thereof
  • Tubercle bacillus recombined protein and expressing purefying method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0052] 1. Construction of recombinant protein Ag85B-Mpt64(190-198)-Mtb8.4:

[0053] 1) First: design primers to amplify mpt64(190-198)-mtb8.4 (8.4f for short),

[0054] Upstream: ACA ATGAGGCTGTCGTTGAC

[0055] Note: The double underline represents the gene encoding the peptide of mpt64190-198.

[0056] Single underline represents: Sal I restriction site

[0057] Downstream: GCG AAGCTT TTAATAGTTGTTGCAGGAG

[0058] Description: Single underline HindIII restriction site

[0059] Using the standard tuberculosis strain (H37Rv) DNA as a template, the above two primers were used for amplification. The PCR reaction conditions were as follows: 98°C pre-denaturation for 5 minutes; 98°C denaturation for 45 seconds, 62°C renaturation for 45 seconds, and 72°C extension 2 minutes and 30 seconds, 30 cycles; extension at 72°C for 12 minutes. After the PCR product was purified, it was digested with Sal I and HindIII, and the clone was constructed in the plasmid PET28a(+).

[0060] 2) Amplify the...

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Abstract

The invention relates to anti-tubercule bacillus recombination protein. It belongs to gene recombination technique field. Its manufacturing includes the following steps: combining antigen the 190-198 polypeptides of the Ag85B-Mpt64 with Mtb8.4; cloning into escherichia coli vector to express; further purifying to gain. It is recorded as Ag85B-Mpt64190-198-Mtb8.4(SEQ.ID.NO.1). The protein can be used as anti tubercule bacillus subunit vaccine.

Description

Technical field [0001] The invention belongs to the technical field of gene recombination, and specifically relates to a Mycobacterium tuberculosis recombinant protein Ag85B-Mpt64 190-198 -Mtb8.4 and its expression and purification method and application. Background technique [0002] Mycobacterium tuberculosis is the pathogenic bacterium that causes tuberculosis in humans and animals. Due to its unique biological characteristics, such as thick cell wall, long growth cycle, mostly latent infection and easy drug resistance, tuberculosis drug treatment is facing great difficulties. Although in the first half of the last century, the Bacille Calmette-Guerin (BCG) vaccine to prevent tuberculosis and specific drugs for the treatment of tuberculosis (such as isoniazid, rifampicin) were successfully developed, by the end of the last century, tuberculosis was still a global infection and contagion. The first "killer" of the disease. Moreover, with the raging AIDS (HIV) worldwide and the ...

Claims

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Application Information

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IPC IPC(8): C12N15/31C12N15/63C12N15/70C12N1/21C07K14/35A61K39/04A61K48/00
Inventor 王洪海郄亚卿祝秉东王九玲徐颖王庆忠
Owner FUDAN UNIV
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