GENETIC ANALYSIS OF NUCLEAR RNA TRANSPORT USING SV40

目前，存在多种SV4O突变体，其表现出与SV4O的表达相关的功能。 复杂的RNA转运表型。 虽然许多这些 突变体破坏RNA剪接功能，通常不转运DNA， 剪接和转运之间存在着明显的关系 因为已知其他突变体转运未剪接的RNA。 我建议 进一步研究RNA剪接和转运之间的关系。 我的具体目标是：1）通过基因操作， SV40基因转录的重要调控元件。 2)为了在体外构建一系列明确的缺失，插入， 和涉及假定的控制区的重排突变体， SV40转录本的剪接和转运过程。 3)以确定 上述突变体的体内转录表型，并建立 这些遗传扰动如何影响剪接、转运和翻译 表达的RNA。 4)为了验证原子核活跃的假设 保留细胞核RNA并确定细胞核与细胞质的区别 核糖核酸 5)解剖修饰的亚核定位 SV40 RNA在剪接和转运过程中的作用。 6)建立 核孔复合体和核基质参与了细胞的增殖， SV40 RNA的剪接和转运，并建立体外转运 系统

Currently there exists a variety of mutants of SV4O which display a complicated range of RNA transport phenotypes. Although many of these mutants perturn RNA splice functions and often do not transport DNA,k no clear relationship has been observed to exist between splicing andtransport as other mutants are known to transport unspliced RNA. I propose to further investigate the relationship between RNA splicing and transport. My specific aims are: 1) to determine, by genetic manipulation, the important control elements that regulate the transcription of SV40genes. 2) To construct,k in vitro, a series of well-defined deletion, insertion, and rearrangement mutants involving putative control regions and may affect the splicing and transport process of SV40 transcripts. 3) To determine the in vivo transcriptional phenotypes of the above mutants and establish how these genetic perturbations affet splicing, transport, and translation of the expressed RNAs. 4) To test the hypothesis that nuclei actively retain nuclear RNA and define what distinguishes nuclear from cytoplasmic RNA. 5) To dissect the subnuclear localization of modifictions undergone by SV40 RNA during the process of splicing and transport. 6) To establish the involvement of the nuclear pore complex and the nuclear matrix in the splicing and transport of SV40 RNA and to establish an in vitro transport system.