A rapid enzymatic DNA sequencing technique: determination of sequence alterations in early simian virus 40 temperature sensitive and deletion mutants.
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.
D000595:Amino Acid Sequence; D000956:Antigens, Viral / Q000235:genetics; D001483:Base Sequence; D002872:Chromosome Deletion; D004256:DNA Polymerase I; D004279:DNA, Viral / Q000235:genetics*; D004926:Escherichia coli / Q000201:enzymology; D008722:Methods; D009154:Mutation; D013539:Simian virus 40 / Q000235:genetics*; D013696:Temperature
D000956:Antigens, Viral; D004279:DNA, Viral; D004256:DNA Polymerase I