Marschall, M. and Motz, M. and Leser, Ulrike and Schwarzmann, F. and Oker, B. and Wolf, Hans J.
Hepatitis B virus surface antigen as a reporter of promoter activity.
Gene 81 (1), pp. 109-117.
The coding sequence for the hepatitis B virus surface antigen (HBsAg) was used as a new reporter gene for studies on eukaryotic promoter activity and upstream regulatory sequences. The data observed in transfection assays were comparable to results obtained with conventional chloramphenicol acetyltransferase (CAT) assays, as was demonstrated using various transcriptional regulation sequences. The expression of HBsAg as a reporter protein offered some advantages: (i) In transient expression assays, a time course of promoter activity depending on variable culture conditions could be monitored over a period of time, since the HBsAg was secreted into the culture supernatant. (ii) Evaluation of HBsAg from supernatant aliquots and quantification of the corresponding promoter activities could be performed easily, using the very sensitive and readily available diagnostic HBsAg kits. (iii) In contrast to the conventional CAT assay, the cells remained available for further tests, e.g., Western blot, immunofluorescence or transcript analysis. Characteristics of several Epstein-Barr virus (EBV) promoters, depending on the virus state of EBV-positive B-cells (latency, chemical induction, lytic superinfection, trans-activation), were assayed using the HBsAg reporter system.
|Institutions:|| Medicine > Lehrstuhl für Medizinische Mikrobiologie und Hygiene|
|Hepatitis B Surface Antigens/immunology||MESH|
|Hepatitis B virus/immunology||MESH|
|Herpesvirus 4, Human/genetics||MESH|
|Promoter Regions, Genetic||MESH|
|Regulatory Sequences, Nucleic Acid||MESH|
|Subjects:||600 Technology > 610 Medical sciences Medicine|
|Refereed:||Yes, this version has been refereed|
|Created at the University of Regensburg:||Yes|
|Deposited On:||13 Apr 2011 09:14|
|Last Modified:||21 Jul 2011 02:11|