
Defined Fragments of Calf, Human, and Rat DNA Produced by Restriction Nucleases
Author(s) -
Philippsen Peter,
Streeck Rolf E.,
Zachau Hans G.
Publication year - 1974
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1974.tb03573.x
Subject(s) - ecori , dna , restriction enzyme , microbiology and biotechnology , biology , nucleotide , gel electrophoresis , chemistry , genetics , gene
DNA of a number of mammalian species was split with restriction nucleases from Hemophilus influenzae Rd (Hin DNAase) and Escherichia coli carrying the resistance transfer factor RI (EcoRI DNAase). The digests were submitted to polyacrylamide gel electrophoresis. A broad distribution of unseparated large DNA fragments was observed in all cases. In addition, a number of sharp bands were seen, some of which were well separated from the bulk of the large DNA fragments. The DNA fragments in the bands were characterized with respect to their size and frequency within the genome. About 10% of the calf DNA digested with Hin DNAase migrated in eight bands. The size of the corresponding DNA fragments ranged from 150 to a few thousand nucleotide pairs and the frequencies from 4000 to 150000. After digestion with EcoRI DNAase three bands were seen. On incubation of human DNA with EcoRI DNAase two fragments were produced with 320 and 640 nucleotide pairs and frequencies of 55000 and 14000, respectively, which correspond to about 1% of the genome. In digestions with Hin DNAase only the broad distribution of unseparated fragments was seen. From rat DNA EcoRI DNAase produced six bands, Hin DNAase two bands, and both nucleases together nine bands. The sizes of the DNA fragments ranged from 30 to 440 nucleotide pairs and their frequencies from 6000 to 5 corresponding to about 3% of the total DNA. The relationship of the defined DNA fragments produced by the action of restriction nucleases to the various families of repeated sequences is discussed.