[Establishment of reverse genetics system for class I NDV08-004 strain]

Bing Du Xue Bao. 2012 Sep;28(5):496-500.
[Article in Chinese]

Abstract

Based on the genomic sequence of NDV08-004 strain (GenBank accession number FJ794269), seven pairs of primers were designed to amplify the genomic fragments by RT-PCR and cloned into pGEM-Teasy vector. The fragments (named A to G) were sub-cloned into transcription vector pOLTV5 according to the universal RE site and the plasmid named NDV08-004-pO which contained the full length cDNA of NDV08-004 strain was constructed. Three helper plasmids (pCI-NP, pCI-P and pCI-L) together with NDV08-004-pO were co-transfected into BSR T7/5 cells, and the transfection supernatant was inoculated into SPF embryonated eggs to rescue the virus. The virus was rescued successfully and identified by HA and RT-PCR and sequencing. The rescue system constructed in this study provided a good foundation for the further related research.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Chick Embryo
  • Genetic Vectors / genetics
  • Molecular Sequence Data
  • Newcastle Disease / virology
  • Newcastle disease virus / genetics*
  • Plasmids
  • Reverse Genetics / methods*