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

I. Seif | Zendy; George Khoury | Zendy; Ravi Dhar | Zendy

Open Access

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

Author(s) -

I. Seif,

George Khoury,

Ravi Dhar

Publication year - 1980

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/8.10.2225

Subject(s) - biology , mutant , dna , simian , enzyme , sequence (biology) , dna sequencing , virus , microbiology and biotechnology , mutation , virology , base sequence , genetics , gene , biochemistry

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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