[Transient expression and identification of gene P and NP of NDV LaSota strain in two different cells]

Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2010 Feb;24(1):62-4.
[Article in Chinese]

Abstract

Objective: To Construction of P and NP genes eukaryotic expression vectors of Newcastle Disease Virus LaSota strain,study its reverse genetics and functional genome of NDV.

Methods: P, NP genes were amplified and cloned into pGEM-T easy vector and then subcloned into pcDNA3.1 (+) expression vector respectively, the recombinant plasmids were named pcDNA3.1 (+)-P and pcDNA3.1 (+)-NP, Recombinant plasmids were transfected into 293 and BHK-21 cells respectively and were detected using IE and Western blot analysis.

Results: Expression of P, NP genes were detected and confirmed by the IE and WB analysis.

Conclusion: The recombinant eukaryotic plasmids pcDNA3. 1(+)-P, pcDNA3.1 (+)-NP were expressed in 293 and BHK-21 cells successfully. This research may be helpful for further study of reverse genetics and functional genome of NDV.

MeSH terms

  • Animals
  • Cell Line
  • Cricetinae
  • Gene Expression*
  • Genetic Vectors / genetics
  • Genetic Vectors / metabolism
  • Humans
  • Newcastle disease virus / genetics*
  • Newcastle disease virus / metabolism
  • Nucleocapsid Proteins
  • Nucleoproteins / genetics*
  • Nucleoproteins / metabolism
  • Phosphoproteins / genetics*
  • Phosphoproteins / metabolism
  • Viral Proteins / genetics*
  • Viral Proteins / metabolism

Substances

  • Nucleocapsid Proteins
  • Nucleoproteins
  • P phosphoprotein, Newcastle disease virus
  • Phosphoproteins
  • Viral Proteins
  • nucleoprotein, Newcastle disease virus