[Construction of pseudorabies virus Ea TK-/gE-/gp63- mutant strain and the study on its biological property].
Using pseudorabies virus Ea strain as material, we inserted LacZ gene expression cassette into gE gene. After blue plaque and plaque purification, a recombinant virus PRVEa TK-/gE-/LacZ+ generated. Utilizing EcoR I site in LacZ gene, digested PRVEa TK-/gE-/LacZ+ genome DNA was cotransfected into PK-15 cells with plasmid pFBBS, then PRVEa TK-/gE-/gp63- generated after plaque purification. Four pairs of primers amplification demonstrated the virus was pure TK-/gE-/gp63- mutant virus. PCR product sequence indicates there were 205 bp deletion in TK gene; 1247 bp deletion in gE,gp63 and intergenic region of PRVEa TK-/gE-/gp63- mutant virus genome DNA. Inoculation to Balb/C mice with PRVEa TK-/gE-/gp63- indicates the virulence is reduced greatly.
Liu
Zhenfei
Z
Key Laboratory of Agricultural Microbiology, Ministry of Agriculture, Huazhong Agricultural University, Wuhan 430070, China.
Chen
Huanchun
H
He
Qigai
Q
Zhou
Fuchun
F
Fang
Liurong
L
D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D004279:DNA, Viral / Q000235:genetics; D017353:Gene Deletion; D011558:Herpesvirus 1, Suid / Q000235:genetics* / Q000472:pathogenicity; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D011557:Pseudorabies / Q000821:virology; D013552:Swine; D013937:Thymidine Kinase / Q000235:genetics*; D014759:Viral Envelope Proteins / Q000235:genetics*; D014774:Virulence
D004279:DNA, Viral; D014759:Viral Envelope Proteins; C089580:glycoprotein E, Suid herpesvirus 1; C042966:pseudorabies virus glycoproteins; D013937:Thymidine Kinase