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Recombinant bifidobacterium for preparing EV (Enterovirus) 71 vaccine as well as preparation method and application of recombinant bifidobacterium

A bifidobacterium, EV71 technology, applied in the field of genetic engineering, to increase the survival period in vitro, prevent EV71 virus infection, and prevent EV71 infection

Inactive Publication Date: 2013-11-06
邓启文 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But research on EV71 has no precedent

Method used

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  • Recombinant bifidobacterium for preparing EV (Enterovirus) 71 vaccine as well as preparation method and application of recombinant bifidobacterium
  • Recombinant bifidobacterium for preparing EV (Enterovirus) 71 vaccine as well as preparation method and application of recombinant bifidobacterium
  • Recombinant bifidobacterium for preparing EV (Enterovirus) 71 vaccine as well as preparation method and application of recombinant bifidobacterium

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Embodiment 1

[0039] Embodiment one: Preparation of recombinant bifidobacteria for EV71 vaccine:

[0040] Such as figure 1 with figure 2 The concrete method of the present invention shown is:

[0041] (1), construction of high-efficiency expression plasmids in bifidobacteria:

[0042] Based on the pBAD / gIIIA vector (purchased from Invitrogen Company) as the backbone, artificially synthesized recombinant E.SOD (the gene sequence of Escherichia coli peroxidase dismutase [E.SOD: the sequence is E. coli Mn-superoxide dismutase gene (GenBank: X03951.1)]) sequence of positive and negative strands (synthesized by Shanghai Yingjun Company), and the synthesized E.SOD positive and negative strands have BstI1071 enzyme cleavage site on both sides (positive strand sequence See sequence table 1), in a PCR machine (ABI9700), perform a cycle of "94°C, 30 seconds; 65°C, 30 seconds; 72°C, 30 seconds" to become a double-stranded E. SOD gene;

[0043] The pBAD / gIII vector was digested with the B...

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Abstract

The invention discloses a recombinant bifidobacterium for preparing an EV (Enterovirus) 71 vaccine as well as a preparation method and application of the recombinant bifidobacterium and belongs to the technical field of genetic engineering. The recombinant bifidobacterium is transformed into a vector containing VP1 protein. The recombinant bifidobacterium is prepared by the following preparation steps of: (1) carrying out enzyme digestion on a recombinant double-stranded E.SOD (Escherichia coli. Superoxide Dismutase) sequence and pBAD (plasma Beam Assisted Deposition) / gIII vector and connection to obtain a pBAD / S vector; (2) inserting a double-stranded EXS-(His)6-VP1 gene into the pBAD vector subjected to double enzyme digestion and transforming Escherichia coli to obtain a positive clone pBAD-VP1 vector; and (3) transforming the pBAD-VP1 vector into bifidobacterium, screening pBAD-VP1 for transforming positive bifidobacterium to obtain the recombinant bifidobacterium for preparing the EV71 vaccine for treatment. The probiotic oral vaccine prepared by the recombinant bifidobacterium disclosed by the invention is free from injection administration and higher in economy and is a new mode for preventing the EV71 virus infection.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering and relates to a recombinant bifidobacterium, in particular to a recombinant bifidobacterium for preparing EV71 vaccine and its preparation method and application. Background technique [0002] Hand, foot and mouth disease is a common infectious disease caused by enterovirus 71 (EV71). It is called "poliomyelitis" in the 21st century. In sporadic cases, the patient's throat secretions, saliva, herpes fluid, and feces all contain the virus, which can be transmitted through close contact with the human digestive tract, respiratory tract, and secretions. There are various transmission routes and there is no effective vaccine for prevention Determined that EV71 virus is easy to cause outbreaks, so the development of effective vaccines is one of the most urgent problems in the prevention and treatment of hand, foot and mouth disease. [0003] EV71 virus is a member of the genus Enterovirid...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/21A61K39/125A61P31/14C12N15/74C12R1/01
Inventor 邓启文余治健曾位森
Owner 邓启文
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