Open AccessSequenzanalyse von Oligonucleotiden mit Hilfe von Micrococcus‐Nuclease
Author(s) -
Feldmann H.
Publication year - 1967
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1967.tb00113.x
Subject(s) - micrococcal nuclease , oligonucleotide , nuclease , nucleotide , restriction enzyme , chemistry , dna , crystallography , biochemistry , gene , nucleosome , histone
Micrococcal nuclease has been used in determination of oligonucleotide sequences. In order to study the mode of splitting of this enzyme in more detail, seven oligonucleotides of known sequences were digested under varying conditions. The splitting patterns obtained from the tetranucleotides CpCpCpG, ApApApG and ApΨpUpI, the pentanucleotides UpCpCpUpG, CpUpUpUpG, and UH 2 pUH 2 pApApG, and the heptanucleotide CpApApCpUpUpG were determined by analysis of the degradation products. The digestions were carried out with a crystalline sample of the enzyme. The results agree with and extend earlier observations with non‐crystalline fractions of micrococcal nuclease. A comparison of the splitting patterns leads to some generalizations: from all oligonucleotides investigated so far, the 3′‐terminal dinucleoside phosphate and the 5′‐terminal dinucleoside diphosphate were released under appropriate conditions. Tetranucleotides were split preferentially in the middle, pentanucleotides as to give mainly the dinucleotides and the middle mononucleotides. The digestion of longer oligonucleotides gave more complicated splitting patterns. The information obtained from the analyses was sufficient to reconstruct the nucleotide sequences in all cases. Micrococcal nuclease digestion as described here should be a generally applicable method for the sequence determination of oligonucleotides.