Porcine epidemic diarrhea virus genetic engineering subunit oral vaccine
A porcine epidemic diarrhea and genetic engineering technology, which is applied in the field of porcine epidemic diarrhea virus genetic engineering subunit oral vaccine, can solve the problems of vaccine absorption, inability to settle in intestinal mucosa, and harmfulness of bacteria and viruses to human body.
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[0014] see figure 1 , figure 2 and image 3 Shown, the preparation method of a kind of porcine epidemic diarrhea virus genetic engineering subunit oral vaccine of the present invention comprises the following steps:
[0015] 1).Synthetic M gene
[0016] Connect the synthetic M gene to the PES vector, carry out enzyme digestion according to the synthetic sequence instructions, clone them into pMD18-T plasmid vectors, transform competent JM109 cells, pick positive colonies, and extract a small amount of plasmids with the OMEGA kit for identification by enzyme digestion positive plasmid (see figure 1 );
[0017] 2). Construction of expression vector
[0018] The M gene fragment containing the correct direction and sequence was excised with restriction endonucleases EcoRI and SalI, recovered and purified from the gel, and then ligated with the vector plasmid pMG36e overnight at 16°C; the ligated products were mixed with competent cells MG1363 (laboratory Save) After mixing ...
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