Construction and identification of recombinant Pseudorabies virus with glycoprotein gene of rabies virus SRV9 strain

Ye Wei Li, Yang Yang, Ye Liu, Jing Long Wang, Cheng Long Sun, Nai Jun Han, Xiang Yi Liu, Rong Liang Hu

Research output: Contribution to journalArticlepeer-review

Abstract

Objective: To construct and identify recombinant Pseudorabies virus (PRV) with glycoprotein (G) gene of rabies virus (RV) SRV9 strain. Methods: The expression cassettes of G gene of SRV9 strain and report gene Lac Z were cloned into transfer vector p8AA, and the constructed recombinant transfer plasmid p8AA-LacZ-G was co-transfected to RHK-21 cells with genome of PRV Rartha-K61 strain. After appearance of CPE, the virus liquid was collected for several cycles of blue plaque screening, and the obtained recombinant virus rPRV-LacZ-G was observed by electron microscopy, identified by PCR and Western blot, and determined for titer. Results: Restriction analysis and sequencing proved that recombinant plasmid p8AA-LacZ-G was constructed correctly. The constructed rPRV-LacZ-G showed typical structure of PRV under electron microscope. The specific band at a length of 1 790 bp was observed by PCR. Western blot showed specific reaction band of rPRV-LacZ-G with monoclonal antibody against G protein of SRV9 strain. The titer of rPRV-LacZ-G was 106.25 TCID 50/ml, which was lower than that of its parental strain (10 7 TCID50/ml). Conclusion: The recombinant PRV with G gene of rabies virus SRV9 strain, i. e. rPRV-LacZ-G was successfully constructed, which laid a foundation of its application to development of rabies vaccine for animal use.

Original languageEnglish (US)
Pages (from-to)536-538+548
JournalChinese Journal of Biologicals
Volume25
Issue number5
StatePublished - May 20 2012
Externally publishedYes

Keywords

  • Glycoprotein
  • Homologous recombination
  • Lac Z gene
  • Pseudorabies virus (PRV)
  • Rabies virus (RV)
  • SRV9 strain

ASJC Scopus subject areas

  • Biotechnology
  • Immunology
  • Biochemistry
  • Applied Microbiology and Biotechnology

Fingerprint

Dive into the research topics of 'Construction and identification of recombinant Pseudorabies virus with glycoprotein gene of rabies virus SRV9 strain'. Together they form a unique fingerprint.

Cite this