Method for preparing actinobacillus pleuropneumoniae (App) bacterial ghost and method for preparing subunit vaccine by loading pasteurella antigen with App bacterial ghost
A subunit vaccine, Pasteurella technology, applied in bacterial antigen components, botanical equipment and methods, biochemical equipment and methods, etc., can solve problems such as poor cross-protection ability, incomplete cleavage, and lack of antigenic epitopes
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[0043] Below in conjunction with specific embodiment the present invention is described in further detail:
[0044] 1 Materials and methods
[0045] 1.1 Materials
[0046] Restriction endonucleases EcoRI, SalI, ApaI and BamHI, PhiX174 phage RF I plasmid, pMC-Express and pBV220 plasmid were purchased from Fermentas MBI Company, USA. The cloning vector pMD18-T and E.coli DH5α were purchased from Dalian TaKaRa Company. Staphylococcus aureus standard strain ATCC25923 and App 1 Shope 4074 strain were purchased from China Veterinary Drug Control Institute.
[0047] 1.2 Method
[0048] 1.2.1 Design and synthesis of primers
[0049] Two pairs of primers were designed according to the coding sequences of phage PhiX174 cleavage gene E (sequence number: 9626372) and Staphylococcus aureus nuclease A gene SN (sequence number: 21281729) in GenBank:
[0050] E1: 5'-CAG GAATTC ATGGTACGCTGGACTTTGTGG-3' (EcoRI)
[0051] E2: 5'-GGC GTC GAC TTACTCCTTCCGCACGTAAT-3'(SalI)
[0052] SN1: ...
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