Molecular Cloning of 99% of Measles Virus Genome, Positive Identification of 5’ End Clones, and Mapping of the L Gene Region

Abstract

Molecular probes specific for paramyxovirus 3’ and 5’ end sequences are essential for studying viral transcription and replication and the role of defective interfering particles in these processes. We have constructed from measles virus genomic RNA an ordered series of overlapping clones containing over 99% of measles virus genetic information. Clones corresponding to the 3’ and 5’ ends of the genome were identified by means of a probe made from ‘capped’ genomic RNA, and their location was confirmed by primer extension experiments. Clones representing the N, P/C, M, F, and H genes were identified by reference to previously characterized clones, and their positions in the genome were defined by restriction analysis and SI nuclease mapping of intercistronic clones. The L gene region was characterized by site-to-site restriction mapping and measured as 6,550 nucleotides in length.