CpG islands detected by self-primed in situ labeling (SPRINS) |
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Authors: | Claus Lindbjerg Andersen Jørn Koch Eigil Kjeldsen |
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Institution: | (1) The Danish Cancer Society, Department of Cytogenetics, Tage Hansens Gade 2, DK-8000 Aarhus C, Denmark, DK;(2) Department of Clinical Genetics, Odense University Hospital, DK-5000 Odense C, Denmark, DK |
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Abstract: | We have studied the distribution and methylation of CpG islands on human chromosomes, using the novel technique of self-primed
in situ labeling (SPRINS). The SPRINS technique is a hybrid of the two techniques primed in situ labeling (PRINS) and nick
translation in situ. SPRINS detects chromosomal DNA breaks, as in nick translation in situ, and not annealed primers, as is
the case in PRINS. We analyzed in situ-generated DNA breaks induced by the restriction enzymes HpaII and MspI. These restriction enzymes enable the detection of chromosomal CpG islands. Both HpaII- and MspI-SPRINS produce a banding pattern resembling R-banding, indicating a higher level of CpG islands in R-positive bands than
in R-negative bands. Our SPRINS banding observations also indicate differences in sequence copy number in the satellites of
homologous acrocentric chromosomes. Furthermore, a comparison of homologous HpaII-SPRINS-banded X chromosomes of females from lymphocyte cultures grown without methotrexate or bromodeoxyuridine revealed
methylation difference between them. The same comparison of homologous X chromosomes from the cell line GM01202D, which has
four X chromosomes, one active and three inactive, revealed the active X chromosome to be hypermethylated.
Received: 5 February 1998; in revised form: 8 May 1998 / Accepted: 11 May 1998 |
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