Investigation of the methylation status around parent-of-origin detectable SNPs in imprinted genes.
Methylation of CpG dinucleotides was investigated in five regions by bisulphite treatment of gDNA, PCR and cloning/sequencing. The gDNA was prepared from peripheral blood, saliva, semen, nails and hair from the head. In gDNA from peripheral blood, three regions were investigated in 16, 23 and 24 individuals, respectively (Fig. 2). In gDNA from other sources, three or five regions were investigated in five individuals (Fig. 3). In many of the sequenced fragments, all the CpG dinucleotides were either methylated or not, which support the idea that the parental origin of an allele may be determined by the methylation status of the allele. However, the methylation of CpG dinucleotides varies across the fragment in some of the sequenced fragments, especially from semen samples, which indicate that it may be difficult to determine the parental origin from some gDNA sources by restriction-enzyme analysis (DMPA method).
Nakayashiki
Nori
N
Department of Legal Medicine, Iwate Medical University School of Medicine, Morioka 020-8505, Japan. nnakaysk@iwate-med.ac.jp
Takamiya
Masataka
M
Shimamoto
Kirito
K
Aoki
Yasuhiro
Y
Hashiyada
Masaki
M
D000483:Alleles; D018899:CpG Islands / Q000235:genetics; D004247:DNA / Q000097:blood / Q000235:genetics / Q000378:metabolism; D019175:DNA Methylation; D005260:Female; D018392:Genomic Imprinting; D006801:Humans; D008297:Male; D020641:Polymorphism, Single Nucleotide; D015183:Restriction Mapping / Q000379:methods; D017422:Sequence Analysis, DNA / Q000379:methods; D013447:Sulfites / Q000494:pharmacology; D055517:snRNP Core Proteins
D013447:Sulfites; D055517:snRNP Core Proteins; D004247:DNA; C042345:hydrogen sulfite