Abstract
This chapter describes a method developed for characterizing and comparing nucleotide sequences in DNA's. The method is based upon the fact that DNases are sequence-specific—that is, they are able to recognize the nucleotides near the phosphodiester bonds that they split. The chapter demonstrates this point for at least the three DNases that have been investigated: spleen acid DNase, snail acid DNase, and pancreatic DNase. As DNases are sequence-specific, the analysis of termini, namely, the determination of the base composition of the termini released from the sequences split by DNases, provides information on the frequency of these sequences in the DNA's analyzed. The analysis of termini is characterized by two features that distinguish it from a determination of the frequency of the sequences split by the enzymes. The analysis of termini formed by DNase is a new method for characterizing and comparing nucleotide sequences in DNA's. When applied to the 3' terminal, the 5' terminal, and the 5' penultimate nucleotides, the method requires 100 μg of DNA. Radioactive labeling of the 3' terminals with [α- 32 P]ATP, using the terminal nucleotidyltransferase, and the 5' ends with polynucleotide kinase should lead to a considerable reduction in scale.
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