Molecular Clones Representing Sendai Virus Genes P, NP and M

Abstract

DNA copies of segments of Sendai virus genes P, NP and M, obtained by reverse transcription of virus mRNA species extracted from chicken embryo lung infected cells, were cloned in plasmid pBR322. Genes were identified by hybrid-arrested translation of viral mRNA in vitro. Hybrid selection of NP mRNA confirmed the identity of an NP gene clone. Partial sequencing of this insert showed that it represents the 5''-terminal region of the gene, containing transcription termination signals. Hybridization of DNA inserts to blots of electrophoretically separated, denatured mRNA species indicated that the P, NP and M messages had sizes of 2400, 2100 and 1500 nucleotides, respectively. Specific T1 RNase-resistant oligonucleotides, previously identified in the NP and M genes, were selected by hybridization to the respective inserts. Although most of the inserts are smaller than 500 base pairs, representing no more than 16% of the P gene and 24% of the NP gene, one of the M gene inserts, comprising 700 base pairs, represents almost half of that gene. These cloned virus gene segments will assist further investigations of the molecular biology of this model paramyxovirus.