Construction of an Eukaryotic Expression Plasmid for gE Gene of Pseudorabies Virus Strain Yue-A

Based on the gE nucleotide sequence of PRV strains NIA3 and Rice, a pair of primers were designed. Using the genome DNA of the strain YueA of PRV , which was isolated from an outbreak in Guangdong Province, as template, a segment of gE was amplified by polymerase chain reaction(PCR), and a fragment with the expected size was obtained. The gE gene fragment was cloned into PMD18T vector, and the recombinant plasmid named PMD18gE obtained. The gE fragment was recovered after the plasmid PMD18gE was digested with HindⅢand BamHⅠ, then was cloned into the mammalian expression vector pcDNA31(+). Finally, a recombinant plasmid named pcDNA31gE, which was identified by PCR, restriction enzyme analysis and sequencing ,was obtained.