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Nucleic Acids Research, 1975, Vol. 2, No. 6 839-852
© 1975

Articles

Determination of the sequences of 18 nucleotides from the 5'-end of the 1-strand and 15 nucleotides from the 5'-end of the r-strand of T7 DNA+

Peter C. Loewen

Department of Microbiology, University of Manitoba Winnipeg, Manitoba, Canada, R3T 2N2

Received March 26, 1975.

The sequences of 18 nucleotides from the 5'-end of the 1-strand and 15 nucleotides from the 5'-end of the {gamma}-strand of T7 bacteriophage DNA have been determined to be pT-C-T-C-A-C-A-G-T-G-T-A-C-G-T-C-C-C (1-strand) and pA-G-G-G-A-C-A-C-A-G-C-G-C-T-C (r-strand)*. The 5'-termini of whole DNA or separated strands were kinased using polynucleotide kinase and ({gamma}-32P; rATP. The DNA was partially digested with pancreatic DNase and the fragments were separated by two dimensional electrophoresis and homochromatography. To complete the sequence, snake venom phosphodiesterase digestions of these fragments were carried out. The relationship of these sequences to the proposed cleavage of concatemeric DNA during DNA replication is discussed.

+Research supported by the National Research Coucil of Canada and the University of Manitoba Faculty of Graduate Studies


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