Peptide mapping characterization of viral proteins generated in a cell-free coupled system for the transcription and translation of influenza virus mRNA

Abstract

In a coupled cell-free system for the transcription and translation of the influenza mRNA's, containing detergent-disrupted purified NWS influenza virion and a micrococcal nuclease-preincubated rabbit reticulocyte lysate, five unglycosylated viral proteins (NS1, M, NP, P1, and P3) were easily produced and isolated. Their identification was based on the electrophoretic separation of peptide fragments resulting from their partial digestion with proteases of restricted specificity (D.W. Cleveland, S. G. Fisher, N. W. Kirschner, and U. K. Laemmli, J. Biol. Chem. 252:1102-1106, 1977).