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Carrier bacterin of attenuated typhoid bacterium of carrying tubercle branch bacillus Ag85B

A technology of Mycobacterium tuberculosis and Salmonella typhi, which is applied in the fields of molecular immunity and infection immunity, and achieves the effect of low preparation cost

Inactive Publication Date: 2007-10-10
WUHAN UNIV
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Problems solved by technology

But so far no DNA vaccine against tuberculosis is more protective than BCG [13]

Method used

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  • Carrier bacterin of attenuated typhoid bacterium of carrying tubercle branch bacillus Ag85B
  • Carrier bacterin of attenuated typhoid bacterium of carrying tubercle branch bacillus Ag85B
  • Carrier bacterin of attenuated typhoid bacterium of carrying tubercle branch bacillus Ag85B

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Embodiment Construction

[0031] The attenuated Salmonella typhi vector vaccine carrying Mycobacterium tuberculosis Ag85B provided by the invention is prepared by a method comprising the following steps:

[0032] 1. Preparation of prokaryotic recombinant expression plasmid pGEX-Ag85B and protein expression identification:

[0033] Use the PCR method to amplify the 121-975 positions of the target gene, that is, the BX842578 gene sequence of Ag85B, first remove the 1-120 coding product and the last 3 stop codon TGA in the BX842578 gene sequence of Ag85B, and then amplify the target gene. Amplification; the used upstream primer p1 and downstream primer p2 of PCR have the nucleotide sequences shown in SEQ ID NO.1 and SEQ ID NO.2 respectively, namely:

[0034] Upstream primer p1: 5'-T GA ATT C AA ATG TTC TCC CGG CCG-3', containing EcoR I restriction site,

[0035] Downstream primer p2: 5'-A AA GCT T TC AGC CGG CGC CTAA-3', containing HindIII restriction site.

[0036] The prokaryotic expression plasm...

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Abstract

An attenuated Salmonella typhi carrier vaccine carrying tubercular mycobacterium Ag85B is prepared through preparing prokaryotic recombinant expression plasmid pGEX-Ag85B, configuring eukaryotic recombinant expression plasmid pVAX1-Ag85B, and preparing said antitubercular vaccine. It can be orally taken.

Description

technical field [0001] The invention belongs to the fields of molecular microbiology, molecular immunity and infection immunity. In particular, molecular cloning is used to design and prepare an attenuated Salmonella typhi vector vaccine carrying Mycobacterium tuberculosis Ag85B. Background technique [0002] Tuberculosis (Tuberculosis, TB) is an infectious disease that seriously endangers human health, caused by Mycobacterium tuberculosis (Mycobacterium tuberculosis, MTB) infection. According to WHO estimates, 1 / 3 of the global population has been infected with Mycobacterium tuberculosis, with 8-10 million new cases each year, of which about 3 million deaths, the number of deaths from tuberculosis is the sum of the deaths from other infectious diseases [1] . China is the second hardest-hit area of ​​tuberculosis in the world except India. At present, the number of tuberculosis cases in my country has increased by more than one million every year, and the annual death toll...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K39/385A61K39/04C12N1/21C12N15/63
CPCY02A50/30
Inventor 章晓联王坤
Owner WUHAN UNIV
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