The isolation of CpG islands from human chromosomal regions 11q13 and Xp22 by segregation of partlymelted molecules.
We isolated fragments containing parts of CpG islands from human chromosomal regions chosen for expected differences in gene density by segregation of partly melted molecules. Restriction fragments of P1 bacteriophage clones covering a region of 11q13 and those of cosmid clones derived from Xp22 were recovered from bands in denaturing gradient gels that were retained following prolonged exposure to electric field. Forty-five independent fragments derived from 11q13 and five from Xp22 were isolated. Nucleotide sequence analysis revealed that 11 of the 45 fragments from 11q13 contained CpG islands including four derived from known genes in 11q13. None of the five fragments derived from Xp22 resembled CpG islands. The number of CpG island fragments obtained was consistent with the expectation based on the number of Not I restriction endonuclease sites present at these regions. Adjustment of parameters in our quasi-theoretical approach to the rate of fragment dissociation improves the discrimination between retention and non-retention. The results support probable identification of CpG island fragments by their reduced rate of strand dissociation when retarded in a denaturing gradient gel.
Shiraishi
M
M
Oncogene Division and Radiobiology Division, National Cancer Center Research Institute, 1-1 Tsukiji 5-chome, Chuo-ku 104-0045, Tokyo, Japan. mshirais@ncc.go.jp
Oates
A J
AJ
Li
X
X
Hosoda
F
F
Ohki
M
M
Alitalo
T
T
Lerman
L S
LS
Sekiya
T
T
D001483:Base Sequence; D002882:Chromosomes, Human, Pair 13; D018899:CpG Islands; D004247:DNA / Q000032:analysis* / Q000302:isolation & purification; D016208:Databases, Factual; D015894:Genome, Human; D006801:Humans; D008969:Molecular Sequence Data; D014960:X Chromosome
D004247:DNA