Identification of specific B cell linear epitopes of mycoplasma hyorhinis P37 protein using monoclonal antibodies against baculovirus-expressed P37 protein

摘要

Identification of recombinant plasmid and shuttle plasmid. The restriction map and primer-binding sites. The P37 gene (1140 bp) was subsequently cloned into the expression vector pFastBac™1 via two restriction sites ( H I and I). Double enzyme digestion ( H I and I) revealed specific bands at 4693 bp and 1140 bp. Lane M indicates the DNA molecular quality standard. Lanes 1, 2, and 3 represent plasmids extracted after single-colony expansion of randomly selected white colonies based on the Bac-to-Bac® Baculovirus Expression System instruction Version D (Invitrogen, Carlsbad, CA, USA). The pFastBac™1-His-P37 shuttle plasmid was identified using M13 primers, and a specific band appeared at 3440 bp, whereas pFastBac1 showed a specific band at 2300 bp. Lane M indicates the DNA molecular quality standard. Lane pFastBac1 indicates the pFastBac™1 plasmid negative control. Lanes 1, 2, and 3 represent plasmids extracted after single-colony expansion of randomly selected white colonies as described above