Open AccessMolecular cloning of the 3'-proximal third of Sendai virus genome.
Author(s) -
Peter C. Dowling,
Colomba Giorgi,
Laurent Roux,
Lyle A. Dethlefsen,
Mark Galantowicz,
Benjamin M. Blumberg,
Daniel Kolakofsky
Publication year - 1983
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.80.17.5213
Subject(s) - sendai virus , genome , biology , gene , recombinant dna , cloning (programming) , rna , genetics , dna , virus , microbiology and biotechnology , molecular cloning , virology , polymerase , complementary dna , computer science , programming language
Portions of the Sendai virus genome were randomly cloned by using virion 50S RNA and calf thymus DNA pentanucleotides as primers. The recombinant clones were probed first with radiolabeled products of an in vitro virion RNA polymerase reaction to locate early message clones and then with a probe from the viral genome 3' end to locate the most 3'-proximal clones. Clones were then ordered from the 3' end of the genome and used to construct a genetic map of the 3'-proximal third of the genome by hybrid-selection of mRNAs. We report that the gene order for this region is 3'-NP - P + C - M-5' and that the genetic loci of the viral P and C proteins cannot be separated by these techniques.