Establishment of reverse genetics system for class I NDV08-004 strain.
- Author:
Yun-Xia CHEN
1
;
Hua-Lei LIU
;
Feng GUAN
;
Dong-Xia ZHENG
;
Yun-Ling ZHAO
;
Zhi-Liang WANG
Author Information
1. China Animal Health and Epidemiology Center, Qingdao 266032, China. chenyunxia2008@163.com
- Publication Type:Journal Article
- MeSH:
Animals;
Base Sequence;
Chick Embryo;
Genetic Vectors;
genetics;
Molecular Sequence Data;
Newcastle Disease;
virology;
Newcastle disease virus;
genetics;
Plasmids;
Reverse Genetics;
methods
- From:
Chinese Journal of Virology
2012;28(5):496-500
- CountryChina
- Language: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.
