Construction and application of subgenomic replicon vectors of Japanese encephalitis virus / 病毒学报
Chinese Journal of Virology
; (6): 33-38, 2007.
Article
in Zh
| WPRIM
| ID: wpr-334913
Responsible library:
WPRO
ABSTRACT
Based on the infectious clone of JEV vaccine SA14-14-2, the subgenomic replicons pCTCJEV, pCTMJEV with large deletions in the structural region were constructed. Then they were transfected into BHK-21 cell, the RNA replication of JEV subgenome can be monitored by RT-PCR and the non-structural protein can be found expressed in the cell by IFA. To explore the possibility of using a reporter gene assay to monitor synthesis of the positive-strand and the negative-strand JEV RNA, we inserted an enhanced green fluorescence protein (EGFP) gene into the 3'-UTR of pCTCJEV, pCTMJEV under the control of the internal ribosomal entry site (IRES) of encephalomyelocarditis virus RNA. After transfection, the EGFP fluorescence could be seen under the fluorescence microscope 1 day later,and maintained for more than a week with no apparent cytopathic effect. The constructed JEV replicons would provide valuable tools to provide a possible vector for a long-lasting RNA virus expression system.
Full text:
1
Database:
WPRIM
Main subject:
Replicon
/
Time Factors
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RNA, Viral
/
Cell Line
/
Promoter Regions, Genetic
/
Reverse Transcriptase Polymerase Chain Reaction
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Green Fluorescent Proteins
/
Encephalitis Virus, Japanese
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Genetic Vectors
/
Genetics
Limits:
Animals
Language:
Zh
Journal:
Chinese Journal of Virology
Year:
2007
Document type:
Article