Canine adenovirus type I subunit vaccine and preparation method thereof
A technology of adenovirus type I, applied in the field of canine adenovirus type I subunit vaccine and its preparation, to achieve the effects of improving proliferation activity, strong immunity, and simple preparation method
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[0040] Example 1 Preparation of canine adenovirus type I penton basal protein subunit vaccine
[0041] (1) Construction of plasmid pColdII-PB: use the genome of CAdV-1 F1301 virus strain as a template, design PB and Knob primers, and use PCR method to amplify PB (1434bp) and Knob (519bp). PCR amplification results are shown in figure 1 .
[0042] Table 1 The sequences of primers P1 and P2
[0043]
[0044] The sequence of table 2 primers Knob-F and Knob-R
[0045]
[0046] After the product was purified, it was digested with SacI and PstI, and the 1434bp fragment was recovered. At the same time, the plasmid pColdII was digested with SacI and PstI, and the 3966 fragment was recovered. The two DNA fragments recovered above were treated with T 4 DNA ligase connection, after the connection product was transformed into competent Escherichia coli DH5α, cultivated on the LB solid medium containing ampicillin, and screened the pColdII-PB and pColdII-Knob containing plasmids (...
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