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PCR Cloning and Subcloning | … Materials and methods. Antigen production. The IVA M2 cytoplasmic domain gene was synthesized by Genscript Inc. The gene fragment included BssH II and Not I restriction enzyme sites, used for cloning and a TEV tag, 29 and … | Get A Quote |
Interactions between the cytoplasmic domains of viral transmembrane proteins and host machinery often determine the outcome of viral infection. The M2 protein of influenza A has been identified as a key player in autophagy-mediated viral replication. Here, we describe the engineering and validation of an antibody specific for the cytoplasmic domain of the M2 protein. Through phage and yeast display selection techniques, we obtained an antibody that recognizes: 1) the M2 cytoplasmic domain purified from bacterial inclusion bodies and refolded, 2) full-length M2 recombinant protein expressed in mammalian cells, and 3) native M2 protein in influenza A infected cells. This antibody can serve as a molecular tool to ... More