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Nucleic Acids Research, 1980, Vol. 8, No. 10 2225-2240
© 1980

Articles

A rapid enzymatic DNA sequencing technique: determination of sequence alterations in early simian virus 40 temperature sensitive and deletion mutants

I. Seif, G. Khoury and R. Dhar

Laboratory of Molecular Virology, National Cancer Institute, National Institutes of Health Bethesda, MD 20205, USA

Received March 3, 1980. We have adapted a rapid sequencing technique from the enzymatic nick-translation method of Maat and Smith. The Forward-Backward procedure employs both synthetic and 3' to 5' exonucleolytic activities of E. coli DNA polymerase I to achieve greater reliability, especially in reading stretches of the same nucleotide. The technique has been employed to determine sequence alterations in four early SV40 temperature-sensitive (tsA) point mutants and five early SV40 viable deletion mutants. The nucleotide sequence of these mutants provides an insight into their biological properties.


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[Abstract] [PDF]


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