he UL24 gene of Equine herpes virus type 1 is conserved across many herpesviruses, but its protein product has not been identified. We expressed the UL24 gene in Glutathione S-transferase (GST) Gene Fusion System. SDS-PAGE analysis revealed that the recombinant protein UL24 (pUL24) was not overexpressed in Escherichia coli BL21 host cells after induction by different IPTG concentrations, temperatures and times. That was attributed to transmembrane cytotoxic effects of the protein. The N terminus containing the first 477 bp was deleted and the remaining C terminus 342 bp was cloned into PGEX-6p-1 for construction of a recombinant plasmid PGEX-6p-1/UL24C. SDS-PAGE analysis revealed that the recombinant protein UL24C (pUL24C) was overexpressed in Escherichia coli BL21 host cells. The UL24 c-terminus protein about 11 KDa named pUL24C was purified and used to immunize guinea pig, producing polyclonal antibody. In immunoblotting experiments, this antiserum recognized a 37KDa protein in lysates from infected cells. The specificity and sensitivity of anti-pUL24C serum were detected with Agar gel immunodiffusion reaction. The pUL24C polyclonal antibody can be used for further characterization concerning the dynamic expression of UL24 protein and intracellular localization of UL24 protein in EHV-1 infected cells. |
