title: [Establishment of reverse genetics system for class I NDV08-004 strain].
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.
authors.affiliation: China Animal Health and Epidemiology Center, Qingdao 266032, China. chenyunxia2008@163.com
authors.forename: Yun-Xia
authors.initials: YX
authors.lastname: Chen
authors.forename: Hua-Lei
authors.initials: HL
authors.lastname: Liu
authors.forename: Feng
authors.initials: F
authors.lastname: Guan
authors.forename: Dong-Xia
authors.initials: DX
authors.lastname: Zheng
authors.forename: Yun-Ling
authors.initials: YL
authors.lastname: Zhao
authors.forename: Zhi-Liang
authors.initials: ZL
authors.lastname: Wang
mesh_terms: D000818:Animals; D001483:Base Sequence; D002642:Chick Embryo; D005822:Genetic Vectors / Q000235:genetics; D008969:Molecular Sequence Data; D009521:Newcastle Disease / Q000821:virology; D009522:Newcastle disease virus / Q000235:genetics*; D010957:Plasmids; D059386:Reverse Genetics / Q000379:methods*