[Construction of pseudorabies virus SH strain with gE-gI gene partial deletion mutant including GFP reporter gene].
On the basis of cloning and indentifying gE-gI gene of pseudurabie virus SH strain, the transfer plasmid vector was constucted in order to get the gE-gI gene partial deletion mutant. At first, gE gene and gI gene were cloned into pUC18, constructed the pgEI vector. Then, the 5' terminal sequence of gE gene was deleted 363bp using the restrict endonuclease in gE gene. The GFP expressing cassette was inserted into the deleting site. The recombinant plasmid pgEI including GFP reporter gene deleted part of gE-gI gene was constructed. BHK-21 cell which was infected with PRV-SH for 1-2h were tansfected with the complex of pgEI-GFP and DOTAPA deletion mutant was selected and purified many times in BHK-21 cell through GFP. Inoculation of mice with 2.0X107 PFU of the recombinant virus revealed that mice were partly protected against challenge with PRV-SH containing 2MLD.
Jiang
Yan
Y
Key Laboratory of Animal Diseases Diagnostic & Immunology of Agricultural Ministry, Nanjing Agricultural University, Nanjing 210095, China. jiangyan1029@yahoo.com.cn
Hou
Yufeng
Y
Chen
Puyan
P
D000818:Animals; D017353:Gene Deletion; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics; D011558:Herpesvirus 1, Suid / Q000235:genetics* / Q000276:immunology / Q000502:physiology; D051379:Mice; D008813:Mice, Inbred ICR; D010957:Plasmids; D011995:Recombination, Genetic; D014759:Viral Envelope Proteins / Q000235:genetics*
D014759:Viral Envelope Proteins; C089580:glycoprotein E, Suid herpesvirus 1; C042966:pseudorabies virus glycoproteins; D049452:Green Fluorescent Proteins