A kind of universal Riemerella anatipestifer yolk antibody and its preparation method and application
A technique for Riemer's anatipestifer, egg yolk antibody, which is applied in the field of universal Riemerella anatipestifer egg-yolk antibody and preparation thereof
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[0033] 1. Construction of pET-28a-Porin vector
[0034] 1. With the genomic DNA of Riemerella anatipestifer ATCC 11845 as template, carry out PCR amplification, the amplified product is 1185bp (as shown in SEQ ID NO.2), and its primer sequence is as follows:
[0035] Porin F-BamHI: 5'-GGATCCGTACCAGATACTATTATCGTTC-3' (SEQ ID NO.3),
[0036] Porin R-Not I: 5'-GCGGCCGCAATTCCTATTTCTACTTGAAATCT-3' (SEQ ID NO. 4).
[0037] The reaction system (total volume 20 μL) is:
[0038]
[0039] The amplification reaction conditions are:
[0040]
[0041] Agarose gel electrophoresis, the results of figure 1 shown. Using Tiangen Agar Gel Recovery Kit (DP130419) to recover the PCR products by gel cutting.
[0042] 2. Ligate the recovered PCR product to the pMD-19T simple vector (this embodiment uses a commercially available ligation kit, such as Treasure Biological pMD TM 19-TVectorCloningKit), transform Escherichia coli DH5α, and extract the recombinant plasmid for sequencing. Spec...
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