NUCLEAR MATRIX STRUCTURE AND DNA REPLICATION

细胞核是所有生物遗传信息的储存库 真核细胞包括人类的真核细胞。尽管取得了长足的进步 在定义主要基因组功能的基本分子性质时 在DNA复制、转录和RNA拼接和处理方面，我们的 了解这些流程是如何在 细胞核的范围是极其有限的。同样，尽管 DNA(核苷酸序列)背后的遗传密码长期以来一直是 打破了我们对DNA进入染色质的组织的理解 而细胞核中的高级结构仍处于萌芽阶段。 然而，几个领域的进展给了我们一个乐观的理由。 关于未来的成功。开发适当的方法来检查 核建筑的三维基础与新的和 灵敏的功能检测系统，如原位荧光 DNA杂交(FISH)和荧光显微镜检测 复制站点是未来的希望之一。我们的实验室有 我研究核基质及其在DNA复制中的作用已有多年 15年。现在基本的基础工作牵涉到核矩阵作为 DNA复制的组织站点已经设置好了，我们现在 将我们的注意力集中在细胞内复制的三维组织上 原子核。通过将这些3-D方法与强大的技术相结合 多维计算机图像分析和FISH，我们正在制作 三维绘制DNA复制位点图的进展使用 11号染色体的一组DNA探针将进一步使我们能够确定 11号染色体DNA序列的三维组织及作图 存在于各个复制站点。未来的主要方向是 本课题的研究内容包括：(1)三维组织结构的研究 细胞核内的DNA复制位点；(2)蛋白质的定位 在三维复制位置；(3)基因序列的三维作图 人类11号染色体的细胞核；(4)基因复制图谱 人类3-D中的11号染色体

The cell nucleus is the repository for the genetic information of all eucaryotic cells including that of man. Despite considerable progress in defining basic molecular properties of the primary genomic functions of DNA replication, transcription, and RNA splicing and processing, our knowledge of how these processes are organized and regulated within the confines of the cell nucleus is extremely limited. Similarly, although the genetic code behind the DNA (the nucleotide sequence) has long been broken, our understanding of the organization of this DNA into chromatin and higher order structures in the cell nucleus is still in its infancy. Progress in several areas, however, gives one reason to be optimistic about future success. Development of appropriate methods to examine the three dimensional basis of nuclear architecture coupled with new and sensitive detection systems for function, such as fluorescence in situ hybridization (FISH) and fluorescence microscopic detection of DNA replication sites are among the hopes of the future. Our laboratory has been studying the nuclear matrix and its role in DNA replication for over 15 years. Now that basic groundwork implicating the nuclear matrix as a site for the organization of DNA replication has been set, we are now focusing our attention on the 3-D organization of replication in the cell nucleus. By combining these 3-D approaches with the powerful techniques of multi-dimensional computer image analysis and FISH, we are making progress in mapping the DNA replication sites in 3-D. Use of a chromosome 11 set of DNA probes will further enable us to determine the DNA sequence organization of chromosome 11 in 3-D and map the sequences present at individual replication sites. The major directions to be pursued in this project include : (1) Study of the 3-D organization of DNA replication sites in the cell nucleus; (2) Localization of proteins at replication sites in 3-D; (3) 3-D mapping of gene sequences in the cell nucleus for human chromosome 11; (4) Map the replication of genes for human chromosome 11 in 3-D.