The distribution of genes on human chromosomes as studied by in situ nick translation.
We have studied the distribution of potentially active genes on human chromosomes, using two methods: DNAse I hypersensitivity and restriction enzyme--nick translation with enzymes sensitive to methylation of CpG doublets. DNAse hypersensitivity is known to be associated with potentially active genes, and, when the reaction is detected by "in situ" nick translation, produces an R-banding pattern. Digestion of chromosomes with HpaII or CfoI, both of which should preferentially cut unmethylated sequences in the CpG islands associated with the majority of genes, also produces R-banding patterns. Deviations are attributable to overdigestion of the chromosomes, leading to extraction of DNA and loss of the specific sites that were to be detected. Contrary to the results of a number of previous workers, we have failed to demonstrate any differences between the DNAse I hypersensitivity or the degree of methylation of the active and inactive X chromosomes in metaphases from females.
de la Torre
J
J
MRC Human Genetics Unit, Western General Hospital, Edinburgh, U.K.
Sumner
A T
AT
Gosalvez
J
J
Stuppia
L
L
D002874:Chromosome Mapping / Q000379:methods*; D004262:DNA Restriction Enzymes; D003851:Deoxyribonucleases; D004303:Dosage Compensation, Genetic; D015870:Gene Expression; D006801:Humans; D017403:In Situ Hybridization; D007621:Karyotyping; D008745:Methylation; D014960:X Chromosome
D003851:Deoxyribonucleases; D004262:DNA Restriction Enzymes