Figure 5

A. Detection of DPV by Dot-ELISA. The preliminary application of the polyclonal antibody against DPV UL46M was the establishment of Dot-ELISA to detect DPV. The result showed stronger positive signal with the liver sample of DPV, while negative with DHV-1, E. coli (O1), SE, RA, P. multocida and normal saline. 1. DPV, 2. DHV-1, 3. E. coli (O1), 4. SE, 5. RA, 6. P. multocida, 7. normal saline (negative). B. Schematic diagram of the UL46 ORF cloned into the pMD18-T cloning vector. The fragment of UL46 digested with Bam HI and Xho I was cloned into cloning vector pMD18-T at 16°C overnight using DNA Ligation Mix to generate recombinant cloning plasmid named pMD18-T/UL46. C. Construction of the recombinant expression plasmid pET32a (+)/UL46. The fragment of UL46 digested with Bam HI and Xho I from pMD18-T/UL46 was cloned into a 6× His-tagged prokaryotic expression vector pET32a(+) at Bam HI and Xho sites and designated it as expression vector pET32a(+)/UL46.