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.