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Study on the genetic information carried in DNA conformation and physical properties

DNA构象和物理性质中携带的遗传信息的研究

基本信息

批准号：
18570165
负责人：
OHYAMA Takashi
金额：
$ 2.53万
依托单位：
Waseda University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18570165/
关键词：
genetic information DNA conformation genome Physical properties of DNA chromatin gene expression

项目摘要

The T20 is a synthetic 180 by bent DNA fragment that mimics a part of left-handed supercoils. T20 can activate transcription by organizing local chromatin structure in HeLa cells. This study investigated whether T20 could activate transcription in the genome of mouse ES cells. T20 was found to activate the adjacent promoter even in the ES cells. Furthermore, the function of T20 was maintained even when the cells were differentiated into hepatocytes. We also investigated the locus of the reporter by using lacO/lacI-GFP transgene detection system. We constructed a tandem-repeat of the lac operator sequences(64 repeats), ligated it to the reporter construct, and integrated the resulting construct into the genome of ES cells. The T20-containing reporter was found located in the periphery of the nucleus.II. Genetic information carried in physical properties of DNAWe made a program that can calculate the flexibility profile of a given DNA. Using it, we investigated the flexibility profiles of all of the recombination hot spots of Saccharomyces cerevisiae chromosome 3. These sites were indicated to be slightly more flexible when compared to the average flexibility of the yeast genome. In addition, we found that the exons are more flexible than the introns. This difference may function as a signal for some proteins to distinguish these two regions.

T20是一个人工合成的180个弯曲的DNA片段，模仿了一部分左撇子超级线圈。T20可以通过组织HeLa细胞中的局部染色质结构来激活转录。本研究探讨了T20是否能激活小鼠ES细胞基因组中的转录。甚至在ES细胞中，T20也能激活邻近的启动子。此外，即使细胞分化为肝细胞，T20的功能也保持不变。并利用LACO/Laci-GFP转基因检测系统对该报告基因进行了定位。我们构建了Lac操纵子序列的串联重复序列(重复序列)，将其连接到报告构建体，并将得到的构建体整合到ES细胞的基因组中。含有T20的报告被发现位于核的外围。II.DNA物理性质中携带的遗传信息我们编制了一个程序，可以计算给定DNA的柔韧性。利用它，我们研究了酿酒酵母3号染色体上所有重组热点的柔韧性。与酵母基因组的平均柔韧性相比，这些位置的柔韧性略高。此外，我们还发现外显子比内含子更灵活。这种差异可能是某些蛋白质区分这两个区域的信号。