Trans Complementation of Replication-defective Omsk Hemorrhagic Fever Virus for Antiviral Study

摘要

complementation of OHFV-ΔNS1 RNA with exogenous expression of NS1. Schematic of the construction of OHFV-ΔNS1 clone. By fusion PCR, aa 4–298 in the NS1 gene were deleted using the full-length infectious clone as a template. Deletion sites are represented by a dotted box. Schematic of the expression plasmid of OHFV-NS1. pBABEpuro retroviral vector and H I/ R I sites were used for clone construction. The cassette includes full-length NS1 with N-terminal 24 aa of E sequence and C-terminal HA-tag. Detection of expression of OHFV-NS1 by Western blotting assay using anti-HA antibody. complementation of OHFV-ΔNS1 with exogenous expression of NS1. BHK21 cells were sequentially transfected with 2 μg pBABEpuro-OHFV-NS1 plasmid and 1 μg OHFV-ΔNS1 RNA, and NS3 expression was analyzed by IFA at different times post-transfection. As a negative control, 1 μg OHFV-ΔNS1 RNA was transfected into BHK21 cells.