Construction and identification of a vector inserted with gene of T7 RNA polymerase.
- Author:
Hong-hui SHEN
1
;
Bing-ke BAI
;
Hao-dong LIU
;
Sheng-dong LUO
;
Yan HU
;
Jun HOU
;
Zhi-jie WANG
;
Wei KONG
;
Yi-dan BAO
;
Pan-yong MAO
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Cercopithecus aethiops; DNA-Directed RNA Polymerases; genetics; metabolism; Enterovirus A, Human; genetics; physiology; Gene Expression; Genetic Engineering; methods; Genetic Vectors; genetics; metabolism; HeLa Cells; Humans; Plasmids; genetics; metabolism; Transfection; Vero Cells; Viral Proteins; genetics; metabolism; Virus Replication
- From:Chinese Journal of Experimental and Clinical Virology 2011;25(2):146-148
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo develop a system to rescue virus by intracellular expression of T7 RNA Polymerase.
METHODSThe gene of T7 RNA Polymerase was amplified and cloned to VR1012 by molecular biological technology. The expression plasmid VR-1a was then identified. VR-1a and EV71 infectious plasmid were co-transfected in Vero cell. CPE was observed and viral gene viral antigen were detected.
RESULTSThe gene of T7 RNA Polymerase was successfully cloned into vector VR1012. Vero cell developed to CPE after being transfected VR-1a and EV71 infectious plasmid. EV71 gene was amplified by RT-PCR from the culture. EV71 antigen was also detected by ELISA.
CONCLUSIONThe method can be used to rescue virus. It could apply to immunologic research of EV71 DNA vaccine.
