Protocol detail

Adenoviral Vector Encoding HBV Genome

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We designed a pENTR4 vector (Thermo Fisher Scientific, Waltham, MA, USA) for cloning into replication-deficient recombinant adenovirus serotype 5 (Thermo Fisher Scientific, USA), which consists of a HBV 1.3-overlength genome [28 (link)] linked to a click beetle green (CBG99) luciferase separated by a P2A linker site derived from the porcine teschovirus. The P2A linker and the CBG99-luciferase were cloned in frame onto the second open reading frame coding for the HBc protein and after the HBV polyA-sequence. Generation and titration of serotype 5 adenoviruses was performed as described before [29 (link)].

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Manske K., Schneider A., Ko C., Knolle P.A., Steiger K., Protzer U, & Wohlleber D. (2021). In Vivo Bioluminescence Imaging of HBV Replicating Hepatocytes Allows for the Monitoring of Anti-Viral Immunity. Viruses, 13(11), 2273.

Publication 2021

pENTR4 vector

Adenovirus Adenoviruses Beetle Genome Luciferase Open frame Polya Porcine teschovirus Protein Reading frame Replication Titration Vector

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Variable analysis

independent variables

Cloning of HBV 1.3-overlength genome, P2A linker, and CBG99-luciferase into pENTR4 vector

dependent variables

Generation and titration of serotype 5 adenoviruses

control variables

Procedures for generation and titration of serotype 5 adenoviruses as described in [29]

controls

Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.

Annotations

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