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

¹Key Laboratory of Agricultural Microbiology, Ministry of Agriculture, Huazhong Agricultural University, Wuhan 430070, China.

Wei Sheng Wu Xue Bao = Acta Microbiologica Sinica

|February 1, 2003

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.

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Abstract:

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