Establishment of Reverse Genetics System for NDV Isolated from Goose
- VernacularTitle:鹅源新城疫病毒拯救体系的建立
- Author:
Shun-Lin HU
;
Yan-Mei ZHANG
;
Qing SUN
;
Yu-Liang LIU
;
Yan-Tao WU
;
Xiu-Fan LIU
;
- Publication Type:Journal Article
- Keywords:
Newcastle disease virus,cDNA,Transfection,Rescue
- From:
Microbiology
1992;0(03):-
- CountryChina
- Language:Chinese
-
Abstract:
Eight fragments were amplified and cloned into pCR2.1 vector with the designed primers.The fragments,amplified with primer Ⅰ to Ⅶ,were subcloned into transcription vector to construct the plasmid pNDVZJI which contained the full-length cDNA of NDV ZJI strain.The eukaryotic expression vector pCI-L was constructed by subcloning the fragments,amplified with the primer Ⅴ,Ⅵ and Ⅷ,into the expression vector pCI-neo.The full-length cDNA clone,pNDVZJI,with three helper plasmids,pCI-NP、pCI-P and pCI-L,were cotranfected into BSR-T7/5 cell expressing T7 RNA polymerase.After inoculation of transfected cell culture into embryonated chicken eggs from specific pathogen free(SPF)flock,The NDV of ZJI strain was rescued successfully,which laid a good foundation for the further related research.
