Nucleotide sequence analysis of DNA

Autor: Ray Wu, Robert A. Bambara, Ernest Jay, Gurdev S. Ghangas
Rok vydání: 1973
Předmět:
Zdroj: Biochemical and Biophysical Research Communications. 54:998-1007
ISSN: 0006-291X
DOI: 10.1016/0006-291x(73)90793-6
Popis: Two procedures have been developed and applied to the determination of the 3′ terminal sequences of λ DNA and φ80 DNA. In the first procedure, each 3′ terminus was specifically labeled with a single 32P-nucleotide. Radioactive oligonucleotides of different lengths were obtained by partial pancreatic deoxyribonuclease digestion. From the characteristic mobilities of these oligonucleotides in two dimensional fractionation systems, the 3′ terminal sequence -ACCCGCG for the r-strand and -GGTTACG for the l-strand of λ DNA have been determined. In the second procedure, approximately six nucleotides were removed from each 3′ terminus with exonuclease III, and they were replaced with radioactive nucleotides by partial repair synthesis. After enzymatic digestion and sequence analysis, the above sequences have been confirmed. The 3′ terminal sequences in φ80 DNA are identical to those in λ DNA at least up to the fifth nucleotide from the 3′ ends.
Databáze: OpenAIRE