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Establishment of a Plasmid-Based Reverse Genetics System for the Cell Culture-Adapted Hepatitis E Virus Genotype 3c Strain 47832c by Scholz, Johannes; Bächlein, Christine; Gadicherla, Ashish K.; Falkenhagen, Alexander; Tausch, Simon H.; Johne, Reimar is a Medicine article available to read on EtoBox.
What is Establishment of a Plasmid-Based Reverse Genetics System for the Cell Culture-Adapted Hepatitis E Virus Genotype 3c Strain 47832c about?
The hepatitis E virus (HEV) causes acute and chronic hepatitis in humans. Investigation of HEV replication is hampered by the lack of broadly applicable, efficient cell culture systems and tools for site-directed mutagenesis of HEV. The cell culture-adapted genotype 3c strain 47832c, which represents a typical genotype predominantly detected in Europe, has previously been used for several basic and applied research studies. Here, a plasmid-based reverse genetics system was developed for this strain, which efficiently rescued the infectious virus without the need for in vitro RNA transcription. The cotransfection of T7 RNA polymerase-expressing BSR/T7 cells with one plasmid encoding the full-length viral genome and two helper plasmids encoding vaccinia virus capping enzymes resulted in the production of infectious HEV, which could be serially passaged on A549/D3 cells. The parental and recombinant virus exhibited similar replication kinetics. A single point mutation creating an additional restriction enzyme site could be successfully introduced into the virus genome of progeny virus, indicating that the system is suitable for site-directed mutagenesis. This system is the first plasm
Who reads Establishment of a Plasmid-Based Reverse Genetics System for the Cell Culture-Adapted Hepatitis E Virus Genotype 3c Strain 47832c?
It is typically read by researchers, students, and practitioners in Medicine.
- Author
- Scholz, Johannes; Bächlein, Christine; Gadicherla, Ashish K.; Falkenhagen, Alexander; Tausch, Simon H.; Johne, Reimar
- Publisher
- MDPI AG; Basel: MDPI AG, 2012- (ISSN 2076-0817)
- Published
- 2020
- Language
- EN
- Field
- Medicine (Life Sciences)