Rapid sequence determination of late simian virus 40 16S mRNA leader by using inhibitors of reverse transcriptase. | Zendy

Minou Bina-Stein | Zendy; Marilyn M. Thorén | Zendy; Norman P. Salzman | Zendy; J A Thomspon | Zendy

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Rapid sequence determination of late simian virus 40 16S mRNA leader by using inhibitors of reverse transcriptase.

Author(s) -

Minou Bina-Stein,

Marilyn M. Thorén,

Norman P. Salzman,

J A Thomspon

Publication year - 1979

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.76.2.731

Subject(s) - reverse transcriptase , messenger rna , biology , microbiology and biotechnology , sequence (biology) , nucleic acid sequence , sanger sequencing , virus , dna , rna , polymerase chain reaction , dna sequencing , virology , gene , genetics

A method for the determination of the primary structure of spliced mRNA junction and leader sequences is described. By analogy to the DNA sequencing procedure of Sanger et al. [Sanger, F., Nicklen, S. & Coulson, A. R. (1977) Proc. Natl. Acad. Sci USA 74, 5463--5467], we use 2",3'-dideoxynucleoside triphosphates as chain-terminating inhibitors of the reverse transcriptase (RNA-dependent DNA polymerase) reaction. By using specific DNA restriction fragments as primers in combination with this technique, we have determined the sequence of the spliced junction between the body and the leader sequence of the 16S late mRNA of simian virus 40. The method described should be of general utility in mapping spliced mRNA regions for which the corresponding protein sequence (if any) is unknown.

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