Insertion of EGFP into the replicase gene of Semliki Forest virus results in a novel, genetically stable marker virus.
J Gen Virol
; 88(Pt 4): 1225-1230, 2007 Apr.
Article
in En
| MEDLINE
| ID: mdl-17374766
ABSTRACT
Alphavirus-based vector and replicon systems have been extensively used experimentally and are likely to be used in human and animal medicine. Whilst marker genes can be inserted easily under the control of a duplicated subgenomic promoter, these constructs are often genetically unstable. Here, a novel alphavirus construct is described in which an enhanced green fluorescent protein (EGFP) marker gene is inserted into the virus replicase open reading frame between nsP3 and nsP4, flanked by nsP2 protease-recognition sites. This construct has correct processing of the replicase polyprotein, produces viable virus and expresses detectable EGFP fluorescence upon infection of cultured cells and cells of the mouse brain. In comparison to parental virus, the marker virus has an approximately 1 h delay in virus RNA and infectious virus production. Passage of the marker virus in vitro and in vivo demonstrates good genetic stability. Insertion of different markers into this novel construct has potential for various applications.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Semliki forest virus
/
RNA-Dependent RNA Polymerase
/
Viral Nonstructural Proteins
/
Green Fluorescent Proteins
Limits:
Animals
Language:
En
Journal:
J Gen Virol
Year:
2007
Document type:
Article
Affiliation country:
Estonia