Nucleotide sequence at the termini of the DNA of Bacillus subtilis phage phi 29.
Phage phi 29 DNA cannot be phosphorylated with polynucleotide kinase and [gamma-32P]ATP because of the presence of a viral protein covalently linked to the 5' termini. The 5' ends can, however, be made susceptible to phosphorylation by treatment with alkali and alkaline phosphatase. Restriction fragments Hpa II C and Hpa II F, corresponding to the right and left ends of phi 29 DNA, respectively, were labeled at the 5' ends with polynucleotide kinase and [gamma-32P]ATP or at the 3' ends with terminal transferase and [alpha-32P]ATP or [alpha-32P]cordycepin 5'-triphosphate. After a secondary cleavage of the labeled fragments, the sequence of the first 150-180 nucleotides at the termini of phi 29 DNA was determined by the method of Maxam and Gilbert. The ends of phi 29 DNA are flush, and a six-nucleotides-long inverted terminal repetition was found. The functional implications of the sequences determined are discussed.
D000469:Alkaline Phosphatase; D001412:Bacillus subtilis / Q000032:analysis*; D001435:Bacteriophages / Q000032:analysis*; D001483:Base Sequence; D004262:DNA Restriction Enzymes; D004279:DNA, Viral; D009838:Oligodeoxyribonucleotides / Q000032:analysis
D004279:DNA, Viral; D009838:Oligodeoxyribonucleotides; D004262:DNA Restriction Enzymes; D000469:Alkaline Phosphatase