[Construction of pseudorabies virus Ea TK-/gE-/gp63- mutant strain and the study on its biological property]

Wei Sheng Wu Xue Bao. 2002 Jun;42(3):370-4.
[Article in Chinese]

Abstract

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.

Publication types

  • English Abstract
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Cells, Cultured
  • DNA, Viral / genetics
  • Gene Deletion*
  • Herpesvirus 1, Suid / genetics*
  • Herpesvirus 1, Suid / pathogenicity
  • Mice
  • Mice, Inbred BALB C
  • Molecular Sequence Data
  • Pseudorabies / virology
  • Swine
  • Thymidine Kinase / genetics*
  • Viral Envelope Proteins / genetics*
  • Virulence

Substances

  • DNA, Viral
  • Viral Envelope Proteins
  • glycoprotein E, Suid herpesvirus 1
  • pseudorabies virus glycoproteins
  • Thymidine Kinase