GENOME DIFFERENCE ANALYSIS BETWEEN MONOZYGOTIC TWINS DISCORDANT FOR NEUROPSYCHIATRIC DISORDERS

神经精神疾病不一致的同卵双胞胎之间的基因组差异分析

基本信息

批准号：
08457250
负责人：
OKAZAKI Yuji
金额：
$ 4.93万
依托单位：
NAGASAKI UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1997
项目状态：
已结题

项目摘要

In 1997, we obtained the following results.1. We applied RLGS to genomes from the monozygotic twins discordant for temporal lobe epilepsy, bipolar disorder and panic disorder. However, we found no reproducible RLGS spot difference between affected and non-affected twins. This year, we found a very rare case of monozygotic twin discordant for autism and collected blood samples with informed consents.2. We tried to cloning DNA fragments included in RLGS spots different between affected and non-affected monozygotic twins with schizophrenia that was found in the experiments last year (Tsujita et al., 1998). We faced with difficulties of the original RLGS method in the cloning of human DNA fragment among a RLGS spot. The reasons why we faced with the cloning problems were that the original RLGS required a large amount of human blood, 80ml, and contained only 0.5 satto moles(5*10^<-19>) DNA in average among a RLGS spot that was 1/10^5 less amount than that of DNA usually available for PCR.Moreover, the DNA fragments digested by the combination of NotI, PvuII, PstI had 5' and 3' overhanging ends that were inconvenient for PCR.Therefore, this year we made an effort to develop a modification of RLGS that included PCR process which increased an amount of DNA contained in a RLGS spot and decreased a required volume of human blood for thc cloning. We have almost completed to develop a modified RLGS and got a couple of RLGS patterns. This new modified RLGS must make the cloning easier if we find discordant spots between identical twins discordant for neuropsychiatric disorders.The method developed by principal investigator that combines genome scanning with discordant monozygotic twins may be available for various kind s of disorders with complex genetic basis in the investigation of genomic mechanisms to be avoided suffering from such disorders by one member of a monozygotic twin pair and not to be avoided by the co-twin.

在1997年，我们获得了以下结果1。我们将RLG应用于颞叶癫痫，躁郁症和恐慌症的单卵双胞胎的基因组。但是，我们发现受影响和受影响的双胞胎之间没有可重现的RLGS斑点差异。今年，我们发现了一个非常罕见的单卵双胞胎对自闭症的不和谐，并通过知情同意收集了血液样本。2。我们试图克隆RLGS斑点中的DNA片段，与去年实验中发现的精神分裂症和非影响的单卵双胞胎不同（Tsujita等，1998）。我们面临着原始RLGS方法在RLGS斑点之间克隆人DNA片段时的困难。我们遇到克隆问题的原因是，原始的RLG需要大量的人体血液，80ml，并且在RLGS位置平均只有0.5个satto moles（5**10^<-19>）DNA，为1 /10^5比通常可用于PCR的DNA的量要少。此外，由Noti，PVUII，PSTI组合消化的DNA片段具有5'和3'的悬垂端，这对PCR带来了不便。努力开发RLG的修饰，其中包括PCR过程，该过程增加了RLGS斑点中包含的DNA量，并减少了所需的人体血液以进行克隆。我们几乎完成了用于开发修改后的RLG并获得了一些RLGS模式。如果我们发现相同的双胞胎之间不一致的神经精神疾病之间的不一致的斑点，则这种新的修改后的RLG必须使克隆变得更加容易。对基因组机制进行的研究是由一对单卵双胞胎对的一名成员避免患有这种疾病的，而不是由二线避免的。