Construction of eukaryotic expression vector of general type in reverse genetic systems of non-segmented negative strand RNA virus.
- Author:
Ying HUANG
1
;
Qing TANG
;
Shou-Feng ZHANG
;
Rong-Liang HU
Author Information
1. State Key Laboratory for Molecular Virology and Genetic Engineering, Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Base Sequence;
Cell Line;
Cloning, Molecular;
Cricetinae;
Eukaryota;
genetics;
Gene Expression;
Genetic Vectors;
chemistry;
genetics;
physiology;
Molecular Sequence Data;
Plasmids;
genetics;
metabolism;
RNA Viruses;
chemistry;
genetics;
physiology;
RNA, Viral;
chemistry;
genetics;
Virus Replication
- From:
Chinese Journal of Virology
2010;26(1):65-70
- CountryChina
- Language:Chinese
-
Abstract:
To construct eukaryotic expression vector of general type in reverse genetics systems of non-segmented negative strand RNA viruses, the multiple cloning site of the plasmid pVAX1 was replaced with HamRz cDNA sequence, a 9 sites linker and HdvRz cDNA sequence through the sequential addition of three adapters, the insertion of which could generate the correct 3' and 5' terminal sequences of the primary viral genomic RNA transcript and facilitate the assembly of the complete viral cDNA sequence. The sequences of the three adaptors were correct after identifying by restriction endonuclease digestions and sequencing. The constructed eukaryotic expression vector could not only be used to assemble viral genome, but also provide the basis for establishing the reverse genetic system rapidly.
