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Apoptosis-promoting recombinant BCG and preparation method thereof

A technology of recombinant BCG and recombinant plasmids, applied in the fields of genetic engineering and tuberculosis vaccine research

Active Publication Date: 2014-10-22
FUDAN UNIV
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Problems solved by technology

Although BCG is effective in preventing tuberculous meningitis and miliary tuberculosis in infants and young children, the protective effect of BCG against tuberculosis in adults is 0-80%

Method used

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  • Apoptosis-promoting recombinant BCG and preparation method thereof
  • Apoptosis-promoting recombinant BCG and preparation method thereof
  • Apoptosis-promoting recombinant BCG and preparation method thereof

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Embodiment Construction

[0033] The present invention will be further elaborated below in conjunction with the accompanying drawings and examples.

[0034] 1. Construction of recombinant BCG

[0035] 1. Construction of recombinant plasmids

[0036] According to the plasmid construction model diagram ( figure 1 ) to construct recombinant plasmids as indicated. According to the data released by the tuberculosis database (http: / / genolist.pasteur.fr / TubercuList / genome.cgi), the base size of the gene ag85b (Rv1886c) encoding Mycobacterium tuberculosis is 978bp (base pair, bp, base pair) , where the 1st to 120bp is the ag85b signal peptide (signal peptide, ASP); according to the data published in GenBank of NCBI (http: / / www.ncbi.nlm.nih.gov), the mRNA base encoding the mouse gene bax The base sequence size is 579bp. Combined with the multiple cloning site (MCS) of the plasmid vector pMV261, forward and reverse primers were designed with Pimer 5 software. The principle of restriction endonuclease sel...

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Abstract

The invention belongs to the technical field of genetic engineering and TB vaccine research, and particularly relates to apoptosis-promoting recombinant BCG and a preparation method thereof. A recombinant plasmid is firstly obtained through sequentially inserting a signal peptide ASP of a TB protective antigen Ag85B and an apoptosis-promoting gene BAX in an escherichia coli-mycobacterium shuttle plasmid pMV261, and the recombinant plasmid is electrically transferred to BCG for recombinant BCG acquisition. Compared with BCG, the recombinant BCG built by the invention rBCG:: ASP-BAX has a strong ability of inducing apoptosis of macrophage, and immunity of the body against the TB mycobacterium can be improved. On the other hand, according to cellular immunity generated when the recombinant BCG induces the body, Th1 serves as the main guide, and the immune protection against the TB mycobacterium is more facilitated. The recombinant BCG rBCG:: ASP-BAX is expected to become a new effective anti-TB vaccine, and has a huge development potential.

Description

technical field [0001] The invention relates to the fields of genetic engineering and tuberculosis vaccine research, in particular to an apoptosis-promoting recombinant BCG and a preparation method thereof. Background technique [0002] One-third of the world's population has been infected with Mycobacterium tuberculosis. Bacillus Calmette-Guerin (BCG) is currently the only vaccine against TB available. Although BCG is effective in preventing tuberculous meningitis and miliary tuberculosis in infants and young children, the protective effect of BCG against tuberculosis in adults is 0-80%. Despite more than 90 years of use of BCG, 1.3 million people die from tuberculosis each year. Moreover, the co-infection of multidrug-resistant tuberculosis (MDR-TB), extensively drug-resistant tuberculosis (XDR-TB) and HIV-TB has made the situation of global tuberculosis prevention and control more severe. [0003] BCG has obvious protective effect on the prevention of tuberculosis in y...

Claims

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Application Information

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IPC IPC(8): A61K39/04A61K48/00C12N15/74C12N15/31C12N1/21A61P31/06A61P37/04
Inventor 王洪海李光华徐颖刘国元宋娜孔聪黄琪
Owner FUDAN UNIV
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