[Construction and identification of a recombinant PRRSV expressing protective antigens of type O foot-and-mouth disease virus]

Sheng Wu Gong Cheng Xue Bao. 2012 Dec;28(12):1431-40.
[Article in Chinese]

Abstract

Using mutation PCR, we cloned the target gene containing 421-480nt (141-160aa) and 598-639nt (200-213aa) of VP1 gene of foot and mouth disease virus (FMDV) into the deleted region (508-532aa) of Nsp2 gene of a highly pathogenic porcine reproductive and respiratory syndrome virus derived vaccine strain (HuN4-F112) that was used as vector. The recombinant cDNA was in vitro transcribed followed by transfection of BHK-21 cells for 36 h. Then, the supernatant of the cell culture was continuously seeded to monolayer of MARC-145 cells for recovery of the recombinant virus. CPE was obviously visible after a couple of passages in the seeded MARC-145, and the rescued virus (designated as rPRRSV-F112-O/VP1ep) was identified by Mlu I digestion, sequencing and immunofluorescence assay. Meanwhile, expression of inserted FMDV epitopes was also detected by indirect immunofluorescence assay with polyclonal antibodies against VP1 protein of FMDV. The analysis of biological characteristics shows that the titer of the rescued recombinant PRRSV (TCID50 = -log10(-6.75)/0.1 mL) was similar to its direct parental virus rHuN4-F112-delta508-532, but higher than rHuN4-F112.

Publication types

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

MeSH terms

  • Animals
  • Antigens, Viral / immunology
  • Base Sequence
  • Capsid Proteins / immunology
  • Cell Line
  • Cysteine Endopeptidases / genetics
  • Epitopes / genetics*
  • Foot-and-Mouth Disease / immunology
  • Foot-and-Mouth Disease / prevention & control
  • Foot-and-Mouth Disease Virus / genetics*
  • Foot-and-Mouth Disease Virus / immunology
  • Molecular Sequence Data
  • Mutation
  • Porcine respiratory and reproductive syndrome virus / genetics*
  • Porcine respiratory and reproductive syndrome virus / immunology
  • Recombination, Genetic
  • Swine
  • Transfection
  • Vaccines, Attenuated / genetics
  • Vaccines, Attenuated / immunology
  • Viral Envelope Proteins / genetics
  • Viral Envelope Proteins / immunology
  • Viral Vaccines* / genetics
  • Viral Vaccines* / immunology

Substances

  • Antigens, Viral
  • Capsid Proteins
  • Epitopes
  • VP1 protein, Foot-and-mouth disease virus
  • Vaccines, Attenuated
  • Viral Envelope Proteins
  • Viral Vaccines
  • Cysteine Endopeptidases
  • nsP2 proteinase