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Recombinant bacillus calmette-guerin vaccine secreting bacteria resuscitation somatomedin and application thereof

A growth-promoting factor and recombinant BCG technology, applied in the field of medicine and biology, can solve the problem of insignificant effect of BCG, and achieve the effect of preventing activation and inhibiting proliferation.

Inactive Publication Date: 2010-10-20
FOURTH MILITARY MEDICAL UNIVERSITY
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Problems solved by technology

The function of Rpf protein is necessary in two cases. One is that when low-density bacteria are inoculated in the culture medium, Rpf is required for reproduction and growth; Rpf is necessary for 10 2 The BCG growth of CFU / ml can obviously promote the growth, but for 10 5 The effect of BCG in CFU / ml is not obvious

Method used

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  • Recombinant bacillus calmette-guerin vaccine secreting bacteria resuscitation somatomedin and application thereof
  • Recombinant bacillus calmette-guerin vaccine secreting bacteria resuscitation somatomedin and application thereof
  • Recombinant bacillus calmette-guerin vaccine secreting bacteria resuscitation somatomedin and application thereof

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Embodiment Construction

[0039] The invention provides a recombinant BCG vaccine that secretes bacterial resuscitation growth-promoting factor. On the basis of constructing exogenous expression vector pDE22-Rpf, it is transfected with BCG to prepare recombinant BCG vaccine, and rBCG-Rpf is obtained through resistance screening; and The expression of the target gene and the immune effect of rBCG-Rpf were verified. The present invention will be further described in detail below in conjunction with the accompanying drawings, which are explanations rather than limitations of the present invention.

[0040] 1. Cloning of Rpf gene

[0041] The Rpf gene was cloned by PCR using M.luteus DNA as a template, and the amplification primer P was designed as follows:

[0042] Upstream primer P1: cga ggatcc a ccatggacac catgact 27;

[0043] Downstream primer P2: tca aagctt t caggcctgcg gcaggac 27;

[0044] The upstream primer P1 introduces a BamHI restriction site ggatcc , the downstream primer P2 introduces...

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Abstract

The invention discloses a recombinant bacillus calmette-guerin vaccine secreting bacteria resuscitation somatomedin and application thereof. The recombinant bacillus calmette-guerin vaccine uses the bacillus calmette-guerin vaccine as host cell. An exogenous expression vector carrying out transfection on the host cell is an expression vector comprising a bacteria resuscitation somatomedin genetic fragment. The recombinant bacillus calmette-guerin vaccine can continuously secrete Rpf factors with bioactivity and can induce a humoral immunity response level and a cellular immune response level. An antibody which is generated by aiming at the exogenous Rpf can interdict the stimulating effect of the Rpf to MTB. The level of inducing IFN-gamma of the recombinant bacillus calmette-guerin vaccine is superior to that of BCG. The recombinant bacillus calmette-guerin vaccine is applied to preparation of a preventive immunity vaccine for MTB infection and / or MTB inapparent infection.

Description

technical field [0001] The invention belongs to the technical field of medicine and biology, and relates to a recombinant BCG vaccine, in particular to a recombinant BCG vaccine that secretes bacteria resuscitating growth-promoting factors and an application thereof. Background technique [0002] 1. Recombinant BCG is an effective way to prepare TB vaccine [0003] Bacillus Calmette-Guerin vaccine (BCG), an attenuated live Mycobacterium bovis, is currently the most effective vaccine against tuberculosis and has been vaccinated by 3.5 billion people worldwide since 1948. With the deepening of research, people found that BCG is not only a live vaccine, but also an expression carrier of foreign genes. This discovery opened up a new field for the application of BCG. [0004] rBCG (Recombinate BCG) uses BCG as an engineering bacterium, uses genetic engineering technology to introduce several foreign genes into the same BCG, and relies on the long-term survival of BCG in the host...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/04A61P31/06
Inventor 师长宏张海袁时芳
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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