Molecular cloning of the genome of poliovirus type 1. | Zendy

Sylvie van der Werf | Zendy; François Brégégère | Zendy; H. Kopecká | Zendy; N Kitamura | Zendy; Paul G. Rothberg | Zendy; P Kourilsky | Zendy; Eckard Wimmer | Zendy; Marc Girard | Zendy

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Molecular cloning of the genome of poliovirus type 1.

Author(s) -

Sylvie van der Werf,

François Brégégère,

H. Kopecká,

N Kitamura,

Paul G. Rothberg,

P Kourilsky,

Eckard Wimmer,

Marc Girard

Publication year - 1981

Publication title -

proceedings of the national academy of sciences

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 5.011

H-Index - 771

eISSN - 1091-6490

pISSN - 0027-8424

DOI - 10.1073/pnas.78.10.5983

Subject(s) - biology , plasmid , complementary dna , pbr322 , microbiology and biotechnology , genome , recombinant dna , poliovirus , cloning vector , rna , restriction enzyme , molecular cloning , rnase h , nucleic acid thermodynamics , rnase p , virology , genetics , dna , gene , virus

Poliovirus cDNA.RNA hybrids were prepared from the Mahoney strain of poliovirus type 1 by using reverse transcriptase (RNA-dependent DNA nucleotidyltransferase) and cloned in the Escherichia coli plasmid pBR322. Bacteria colonies carrying recombinant plasmids were selected by in situ hybridization with virus-specific RNase T1-resistant oligonucleotides. Analysis of the cDNA inserts by restriction mapping and electron microscopy showed that the cloned cDNAs, the longest of which was 3.2 kilobase pairs, originated from various parts of the viral RNA, covering at least 99% of the genome length. Due to overlapping of the clones, the restriction map of the poliovirus genome could be reconstructed. The complete 5' end of the genome was successfully cloned in at least one of the recombinant plasmids, pPV1-366.

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