[Construction of minigenome of Newcastle disease virus of goose origin and its preliminary application].
On the base of obtaining the full length genome sequence of a Newcastle disease virus (NDV) isolated from goose, the minigenome was constructed by replacing all the encoding region with the reporter gene of enhanced green fluorescent protein (eGFP), except the virus regulating sequences relating to replication, transcription and packing of virus genome. The reporter gene could be expressed after it was transfected into the HEp-2 cells infected with helper virus of NDV. This result indicated that the minigenome could be translated by the NDV NP, P and L proteins provided by helper virus. Furthermore, the support plasmids expressing NDV NP, P and L protein were constructed respectively and the function of these plasmids was identified using the minigenome. Additionally, the virus rescue system was optimized by changing the infection dose of the recombinant vaccinia virus expressing T7 RNA polymerase. The work mentioned above will accelerate greatly the rescue of NDV and other relative research.
Zhang
Yan-mei
YM
Infectious Disease Laboratory of Chinese Ministry of Agriculture, Yangzhou University, Yangzhou 225009, China. yanmeizhang2003@yahoo.com.cn
Liu
Yu-liang
YL
Huang
Yong
Y
Jia
Li-jun
LJ
Liu
Xiu-fan
XF
D000818:Animals; D002460:Cell Line; D002642:Chick Embryo; D005777:Geese / Q000821:virology; D017930:Genes, Reporter; D016679:Genome, Viral; D049452:Green Fluorescent Proteins / Q000235:genetics; D006378:Helper Viruses / Q000235:genetics; D009522:Newcastle disease virus / Q000235:genetics*; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins / Q000235:genetics*; D010750:Phosphoproteins / Q000235:genetics*; D010957:Plasmids; D014162:Transfection; D014616:Vaccinia virus; D014764:Viral Proteins / Q000235:genetics*
D019590:Nucleocapsid Proteins; D009698:Nucleoproteins; C055691:P phosphoprotein, Newcastle disease virus; D010750:Phosphoproteins; D014764:Viral Proteins; C409209:nucleoprotein, Newcastle disease virus; D049452:Green Fluorescent Proteins