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行政院國家科學委員會專題研究計畫期中進度報告:雞之視覺剝奪性近視的基因表現及蛋白體研究(1/3)
Date Issued
2003
Date
2003
Author(s)
楊長豪
DOI
912314B002406
Abstract
The prevention and treatment of myopia are important issue of public health in many
countries, especially in Taiwan where the prevalence rate of myopia is extremely high.
The most important complication of extreme myopia is retinal degeneration affecting
the posterior pole that is associated with elongation of ocular axial length.
Unfortunately, the actual mechanism of the development of myopia is still unknown.
Isolation and identification of genes expressed in form deprived ocular tissue may aid
in the molecular evaluation and understanding of possible mechanisms involved in
form deprivation myopia. Subtraction-hybridization PCR method (Fig. 1) is used to
selectively amplify target cDNA fragments and simultaneously suppress nontarget
DNA amplification. It can achieve greater than 1000-fold enrichment of differentially
expressed cDNAs (ie. cDNA from myopic and control eyes). The basic idea of
subtraction-hybridization PCR is those tracers DNA (in our experiment, the cDNA
from myopic eyes) will primarily reassociate with excess driver DNA (in our
experiment, the cDNA from control eyes) while target sequences having no
counterparts in driver will inevitably reassociate with each other, or remain
single-stranded. The reassociated fragments common for driver and tracer are
discarded, and the remaining DNA enriched in target sequences is cloned and
analyzed. With this method, Ishibashi et al. showed the upreglation of crystalline
mRNAs in form-deprived chick eyes (Exp Eye Res 2000 70:153-158). It is
proven that subtraction-hybridization PCR is a powerful tool to study chick myopia.
In this experiment, subtraction hybridization technique was used to evaluated the
differential gene expression in the chick sclera of myopic and control eyes.
countries, especially in Taiwan where the prevalence rate of myopia is extremely high.
The most important complication of extreme myopia is retinal degeneration affecting
the posterior pole that is associated with elongation of ocular axial length.
Unfortunately, the actual mechanism of the development of myopia is still unknown.
Isolation and identification of genes expressed in form deprived ocular tissue may aid
in the molecular evaluation and understanding of possible mechanisms involved in
form deprivation myopia. Subtraction-hybridization PCR method (Fig. 1) is used to
selectively amplify target cDNA fragments and simultaneously suppress nontarget
DNA amplification. It can achieve greater than 1000-fold enrichment of differentially
expressed cDNAs (ie. cDNA from myopic and control eyes). The basic idea of
subtraction-hybridization PCR is those tracers DNA (in our experiment, the cDNA
from myopic eyes) will primarily reassociate with excess driver DNA (in our
experiment, the cDNA from control eyes) while target sequences having no
counterparts in driver will inevitably reassociate with each other, or remain
single-stranded. The reassociated fragments common for driver and tracer are
discarded, and the remaining DNA enriched in target sequences is cloned and
analyzed. With this method, Ishibashi et al. showed the upreglation of crystalline
mRNAs in form-deprived chick eyes (Exp Eye Res 2000 70:153-158). It is
proven that subtraction-hybridization PCR is a powerful tool to study chick myopia.
In this experiment, subtraction hybridization technique was used to evaluated the
differential gene expression in the chick sclera of myopic and control eyes.
SDGs
Publisher
臺北市:國立臺灣大學醫學院眼科
Coverage
計畫年度:91;起迄日期:2002-08-01/2003-07-31
Type
journal article
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