Mechanisms of in situ nick translation of chromosomes using restriction endonucleases.
In situ nick translation of mammalian chromosomes by restriction endonuclease treatment to nick the chromosomal DNA, and 'translation' in the presence of DNA polymerase I and biotinylated dUTP, results in a distinct banding pattern. Further experiments have elucidated the mechanisms producing these bands. The hypothesis is presented that differences in the local conformation of the DNA-protein complex, rather than the DNA sequence itself, lead to the nick translation bands. The different DNase I sensitivity along the chromosomes suggests that the bands, which were clearly evident, reflect morphological units closely related to biological functions.
D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D002875:Chromosomes / Q000502:physiology*; D004262:DNA Restriction Enzymes; D003850:Deoxyribonuclease I; D006801:Humans; D007621:Karyotyping; D008214:Lymphocytes / Q000166:cytology; D014176:Protein Biosynthesis
D004262:DNA Restriction Enzymes; D003850:Deoxyribonuclease I