Isolation And Identification Of Pseudorabies Virus La Strain And Nucleotide Acid Comparison Of Envelope Glycoprotein Gene E

Pseudorabies virus(PRV) causes the pseudorabies(Aujeszky"s disease) in swine. The diseas is characterized by an acute and often fatal infection in piglets and by a variety of clinical signs disease in older pigs, including edcephalitis, pneumonia, increased susceptibility to other respiratory pathogens and abortion. The paper aims at a virus strain isolated in a farm suspected of Psedorabies disease in Shandong. The virus was continually inoculated onto BHK-21 cells and induced typical cytopathogenic effect (CPE). The titers of the virus in BHK-21 cells attained 10"7'46/0. 1ml. After the selection of phagocyte, we gain the rarefied virus clone. Culturing the virus on RK, PK, ST, Vero, Marcl45 and CEF cells, the typical CPE could be seen. All of which conform to the biological the properties of the alphaherpesvirinae and coincide with the control virus (PRV Shanghai strain). The typical Pseudorabies virus particles could be found under electro microscope. The isolated virus was sensitive to chloroform and aether, it was inactivated for 30 min at 56癈. The isolate strain can be neutralized by the positive serum to Pseudorabies virus. 4 rabbits and 8 BALB/C mice were injected with the isolates and the typical clinical syndrome could be observed. The isolate virus designated as PRV La was identified as PRV on the base of the above findings.The DNA extracted from the isolated virus was subjected to PCR analysis utilizing a set of oligonucleotide primers and the sequence have 1757 bases constituting the gene coding the envelope glycoprotein E. the gE gene was cloned into pEGFP-Cl plasmid for the expression in the mammalian cells. The nucleotides of the gE gene were sequenced. An analysis of the ORF and its deduced amino acid sequence were performed with DNAStar Programs. The results indicate that the region of gE gene include one ORF of 1740bp and code a protein of 579 amino acids. Nucleotide acid and amino acidcomparison of envelope glycoprotein gene E utilizing MegAlign program. gE gene of PRV La strain shares highly conserved regions with PRY Ea strain, SH strain-. Fa strains-. MinA strains and Rice strains. The homology of deduced amino acids of PRV La strain with above 5 strains is 98. 1%, 97.4%, 96.6%, 96. 4% and 95. 0% respectively. The differentiation of PRV gE sequence shows regular regionally.