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Generation of a KSHV K13 Deletion Mutant for vFLIP Function Study.

Generation of a KSHV K13 Deletion Mutant for vFLIP Function Study.
Author Information (click to view)

Wang F, Guo Y, Li W, Lu C, Yan Q,


Wang F, Guo Y, Li W, Lu C, Yan Q, (click to view)

Wang F, Guo Y, Li W, Lu C, Yan Q,

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Journal of medical virology 2017 12 15() doi 10.1002/jmv.25009
Abstract

Kaposi’s sarcoma-associated herpesvirus (KSHV)-encoded viral Fas-associated death domain-like IL-1-converting enzyme inhibitory protein (vFLIP) is one of the latently expressed genes and plays a key role in cell survival and maintenance of latent infection by activating the NF-κB pathway. To obtain a genetic system for studying KSHV vFLIP mutation in the context of the viral genome, we generated recombinant viruses lacking the coding sequence (CDS) of vFLIP gene (K13/ORF71) by bacterial artificial chromosome (BAC) technology and the Escherichia coli Red recombination system. After a series of verification with PCR, restriction digestion and sequencing, the K13 deletion bacmids was transfected into a stable viral producer cell line based on iSLK cells to create vFLIP-knockout mutant. Importantly, human umbilical vein endothelial cells (HUVECs) could be de novo infected by vFLIP mutant virus, which are now available for studying the roles of vFLIP in regulation of other KSHV genes and viral pathogenesis. This article is protected by copyright. All rights reserved.

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