Luciferase-Labeled MCMV Infection Quantification

MCMV-Δm157-luc was constructed based on the MCMV-BAC pSM3fr-MCK-2fl [60 (link)] (kindly provided by B. Adler, Munich, Germany). Deletion of the m157 ORF and insertion of the luc sequence into the m157 locus were performed exactly as described [61 (link)]. To quantify organ virus titers, organs were homogenized and virus-tissue suspensions were titrated on a stable mouse embryonic fibroblast cell line. 24 h post titration cells were lysed in luciferase buffer (Roche) and luciferase activity was measured (TriStar²; Berthold Technologies). Virus titer was calculated utilizing a virus standard curve.

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Rattay S., Graf D., Kislat A., Homey B., Herebian D., Häussinger D., Hengel H., Zimmermann A, & Schupp A.K. (2018). Anti-inflammatory consequences of bile acid accumulation in virus-infected bile duct ligated mice. PLoS ONE, 13(6), e0199863.

Publication 2018

TriStar2

Buffer Cell line Cells Deletion Embryonic Fibroblast Insertion sequence Luciferase Mouse Sequence insertion Tissue Titration Virus

Corresponding Organization : University of Freiburg

Top 5 similar protocols

Variable analysis

independent variables

Deletion of the m157 ORF
Insertion of the luc sequence into the m157 locus

dependent variables

Organ virus titers
Luciferase activity

control variables

Stable mouse embryonic fibroblast cell line

controls

Virus standard curve

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