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KMID : 1132720040020010030
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Genomics & Informatics
2004 Volume.2 No. 1 p.30 ~ p.35
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The Study of X Chromosome Inactivation Mechanism in Klinefelter’s Syndrome by cDNA Microarray Experiment
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Jeong Yu-Mi
Chung In-Hyuk
Park Jung-Hoon
Lee Sook-Hwan
Chung Tae-Gyu
Kim Yong-Sung
Kim Nam-Soon
Yoo Hyang-Sook
Lee Su-Man
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Abstract
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To investigate the XIST gene expression and its effect in a Klinefelter’’s patient, we used Klinefelter’’s syndrome (XXY) patient with azoospermia and also used a normal male (XY) and a normal female (XX) as the control, We were performed cytogenetic analysis, Y chromosomal microdeletion assay (Yq), semi-quantitative RT-PCR, and the Northern blot for Klinefelter’’s syndrome (KS) patient, a female and a male control, We extracted total RNA from the KS patient, and from the normal cells of the female and male control subjects using the RNA prep kit (Qiagen), cDNA microarray contained 218 human X chromosome-specific genes was fabricated. Each total RNA was reverse transcribed to the first strand cDNA and was labeled with Cy-3 and Cy-5 fluorescein, The microarray was scanned by ScanArray 4000XL system. XIST transcripts were detected from the Klinefelters patient and the female by RT-PCR and Northern blot analysis, but not from the normal male, In the cDNA microarray experiment, we found 24 genes and 14 genes are highly expressed in KS more than the normal male and females, respectively. We concluded that highly expressed genes in KS may be a resulted of the abnormal X inactivation mechanism.
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KEYWORD
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X Inactivation, XIST, Klinefelter’s syndrome, cDNA chip
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