Support plasmids and support proteins required for recovery of recombinant respiratory syncytial virus

Abstract

Respiratory syncytial virus (RSV) can be recovered from plasmids that separately encode antigenomic RNA and the N, P, L, and M2-1 proteins of the nucleocapsid. However, in a recent study the inclusion of a separate M2-1 expression plasmid was found to be unnecessary (H. Jin, D. Clarke, H. Zhou, X. Cheng, K. Coelingh, M. Bryant, and S. Li, Virology 1998, 251, 206-214). This suggested that the M2-1 protein, which is a transcription antitermination factor, is not required to reconstitute the minimum unit of infectivity, namely a nucleocapsid fully functional for viral transcription and RNA replication. Here we show that the antigenomic plasmid is remarkably efficient as a substitute for an M2-1 expression plasmid in supporting processive transcription by an RSV minigenome. Thus, the simple expedient of omitting an expression plasmid is invalid for evaluating recovery requirements. The issue of the requirement of M2-1 for the recovery of infectious RSV is discussed.