Visualization of replicating genome of hepatitis B virus: development of the method for alpha-Cre complementation and frame overlap

2015 Fiscal Year Final Research Report

• Project/Area Number: 26670221
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Virology
• Research Institution: The University of Tokyo
• Principal Investigator: Saito Izumu 東京大学, 医科学研究所, 教授 (70158913)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Keywords: B型肝炎ウイルス / split Cre / α相補 / 蛍光スクリーニング

Outline of Final Research Achievements

We constructed genomes of hepatitis B virus (HBV), in which PreS region were replaced by split Cre’s (α-Cre andβ-Cre for first and latter halves, respectively), while overlapping Pol frame is maintained. Simultaneous expression of both the split Cre’s resulted in the Cre activity. The replacement does not exceed the genome size that can be replicated. The HBV genome described here expressed split Cre and, when the other split Cre was supplied from outside the genome, functional Cre activity was observed and turn on the GFP expression present outside. Because the replication of HBV genomes constructed here have not sufficiently been examined within the term of this study, further experiments are needed to confirm replication of these genomes quantitatively. Moreover, more information could be available if adenovirus vector systems are used instead of transfection. When they are confirmed, these HBV genomes must be useful for fluorescent screening of ant-HBV medicine.

Free Research Field

ウイルス学