Sequence organisation in nuclear DNA from Physarum polycephalum: methylation of repetitive sequences.
Nuclear DNA from the slime mould Physarum polycephalum is digested by the restriction endonuclease HpaII to generate a high molecular weight and a low molecular weight component. These are referred to as the M+ and the M- compartment, respectively. Sequences that are present in the M+ compartment are cleaved by MspI, the restriction enzyme isoschizomer of HpaII, thus showing that the recognition sequences for these enzymes in M+ DNA contain methylated CpG doublets. The distribution of repetitive sequences in the M+ and M- DNA compartments was investigated by comparison of the 'fingerprint' patterns of total Physarum DNA and isolated M+ DNA after digestion using different restriction endonucleases, and by probing for the presence of specific repetitive sequences in Southern blots of M+ and M- DNA by the use of cloned DNA segments. Both types of experiment indicate that many repetitive sequences are shared by both compartments, though some repetitive sequences appear to be considerably enriched, or are present exclusively, either in M+ DNA or in M- DNA.
D001483:Base Sequence; D002467:Cell Nucleus / Q000032:analysis; D002499:Centrifugation, Density Gradient; D004247:DNA / Q000032:analysis*; D004262:DNA Restriction Enzymes / Q000378:metabolism; D008745:Methylation; D008970:Molecular Weight; D009693:Nucleic Acid Hybridization; D010804:Physarum / Q000235:genetics*; D012091:Repetitive Sequences, Nucleic Acid / Q000187:drug effects*
D004247:DNA; D004262:DNA Restriction Enzymes