2.1 Plasmid construction
Construction of the pCIgD plasmid, which expresses the secreted form of
BoHV1 glycoprotein D (gD), has been previously described (Langellotti et
al., 2011). pCIgD and pCIneo empty plasmid (Promega negative control)
were amplified in transformed Escherichia coli DH5α and purified
using anion exchange columns (Qiagen Plasmid Purification GIGA Kit).
They were analyzed based on 260/280 absorbance ratios and restriction
digests. Integrity and concentration of the plasmids were evaluated
before and after each vaccine formulation.