Amplification, Cioning And Sequence Analysis Of GE Gene Of Pseudorabies Virus B And W Strains

In this study, two fragments of 985 bp and 934 bp were amplified by PCR technique from the DNA genome of PRV Guangxi B and W wild strains according to the published gE gene of PRV Rice strain. These PCR products were purified by Agrose gel ,cloned into PMD18-T Vector and transformed into E.coli DH5 a . Four positive recombinants (PMD18-T GXBi, W, andPMD18-T GXB2, W2) were obtained. The recombinants were screened by PCR and digested by restriction endonucleases Hindlll and EcoRJ. Then the inserts fragments were sequenced. The results revealed that the gE genes were both composed of 1743 base pairs and coded for 580 amino acids. The comparison of homology of the gE genes showed that the homology of nucleotide sequences were 99% , 99.1% homology of deduced amino acids sequence was evaluated between PRV-GXB strain and PRV-GXW strain. The homology of nucleotide sequence between B, W strain and Rice strain were 98.0% and 97.6% respectively and the homology of amino acids were 95.7% and 95.2%. The same work is done on the comparison between B, W strains and several domestic isolated strains. The homology of nucleotide sequence between B, W strain and domestic isolated strains were 98.4%~99.5% and the homology of amino acids were 97.3%~ 99.1%. Phylogenetic tree analysis showed there was a high relationship between GXB, W strains and Ea strain. This study provides scientific theoretical information for the molecular epidemiology of Pseudorabies in Guangxi and lays a good foundation for constructing gE gene delected vaccine and the diagnostic method of identifying PRV.