Analysis of the distribution of CAG repeats and X-chromosome inactivation status of HUMARA gene in healthy female subjects using improved fluorescence-based assay

Citation
M. Karasawa et al., Analysis of the distribution of CAG repeats and X-chromosome inactivation status of HUMARA gene in healthy female subjects using improved fluorescence-based assay, INT J HEMAT, 74(3), 2001, pp. 281-286
Citations number
21
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Hematology
Journal title
INTERNATIONAL JOURNAL OF HEMATOLOGY
ISSN journal
0925-5710 → ACNP
Volume
74
Issue
3
Year of publication
2001
Pages
281 - 286
Database
ISI
SICI code
0925-5710(200110)74:3<281:AOTDOC>2.0.ZU;2-L
Abstract
We investigated the polymorphic CAG-repeat distribution and the X-inactivat ion status of the human androgen receptor (HUMARA) gene in 58 female Japane se volunteers. Polymerase chain reaction amplification was performed using a fluorescent-dye-labeled primer under conditions specific for GC-rich targ ets, and fragments were analyzed. To estimate the length of these fragments , FAM-labeled (blue fluorescent) products were simultaneously compared with ROM-labeled size markers (red) that were created by sequencing various HUM ARA fragments. The number of polymorphic CAG repeats of HUMARA in 116 allel es from 58 female subjects ranged from 15 to 28. Of the 58 volunteers, 51 ( 88.0%) were heterozygous. In 96% of the heterozygous female subjects, the a llelic differences were no greater than 6 repeats. X-chromosome inactivatio n was calculated as the ratio of the area of the smaller peak to the sum of the areas of the smaller and larger peaks. The average ratio was 0.38 (ran ge. 0.09-0.50). Preferential use of 1 allele, by more than 75% (ratio, <0.2 5), was observed in 5 volunteers (10.9%). The clonal nature of a patient wi th chronic myelogenous leukemia was easily identified. This method is sensi tive enough to discriminate a difference of 1 triplet repeat. (C) 2001 The Japanese Society of Hematology.