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Insertion of EGFP into the replicase gene of Semliki Forest virus results in a novel, genetically stable marker virus

Title: Insertion of EGFP into the replicase gene of Semliki Forest virus results in a novel, genetically stable marker virus
Authors: Tamberg, N; Lulla, V; Fragkoudis, R; Lulla, A; Fazakerley, JK; Merits, A
Publisher Information: SOC GENERAL MICROBIOLOGY
Publication Year: 2007
Collection: The University of Melbourne: Digital Repository
Description: 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.
Document Type: article in journal/newspaper
Language: English
ISSN: 0022-1317
Relation: https://hdl.handle.net/11343/261001
Availability: https://hdl.handle.net/11343/261001
Rights: https://creativecommons.org/licenses/by/4.0 ; CC BY
Accession Number: edsbas.2E9BA51A
Database: BASE