Expression of the newcastle disease virus fusion glycoprotein in vero cells using attenuated Salmonella typhimurium as transgenic carrier

Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai). 2002 Jul;34(4):488-93.

Abstract

The full-length cDNA of fusion protein (F) gene of newcastle disease virus (NDV)strain F48E9 was amplified by RT-PCR and inserted into pcDNA3 under the control of human cytomegalovirus (hCMV) immediate early enhancer and promoter. The resulting recombinant plasmid pcDNA3-F was transformed by electroporation into attenuated Salmonella typhimurium strain ZJ111 (dam(-) and phoP(-)), which was then used to transfect the Vero cells. The DNA and RNA dot blotting revealed that the F gene was transcribed into mRNA in the Vero cells. There was expression of the F protein as shown by indirect immunofluorescent assay. The expression began at 48 h post-infection and increased thereafter, as indicated by ELISA. A 55 kD band of the F protein was identified by SDS-PAGE and Western blotting. These results clearly show that the expressed fusion protein was immuno-reactive with chicken anti-NDV serum.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Blotting, Western
  • Chlorocebus aethiops
  • DNA, Complementary / genetics
  • Gene Expression
  • Gene Transfer Techniques*
  • Molecular Sequence Data
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Salmonella typhimurium / genetics*
  • Vero Cells
  • Viral Fusion Proteins / genetics*
  • Viral Fusion Proteins / metabolism

Substances

  • DNA, Complementary
  • RNA, Messenger
  • Recombinant Fusion Proteins
  • Viral Fusion Proteins