Recombinant bifidobacterium for preparing EV (Enterovirus) 71 vaccine as well as preparation method and application of recombinant bifidobacterium
A bifidobacteria, EV71 technology, applied in the field of genetic engineering, to achieve the effects of preventing EV71 virus infection, preventing EV71 infection, and increasing in vitro survival period
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[0039] Embodiment one: Preparation of recombinant bifidobacteria for EV71 vaccine:
[0040] Such as figure 1 and 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 Bs...
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