E. coli Termination Factor Rho, pBR322 Rop, and mRNA Degradation

9612367格罗诺斯塔斯基核因子I(NFI)是一类系统发育保守的位点特异性DNA结合蛋白，在包括生物体发育在内的基本细胞过程中发挥着重要作用。这些研究考察了含有单一NFI基因的有机体中的一个简单的发育系统。NFI在线虫生长发育中的作用将通过利用TCL靶向突变产生NFI基因的缺失突变来研究。纯合子零突变和表达特定突变形式的生物的表型将为这一进化保守的DNA结合蛋白家族在线虫和其他生物中如何发挥作用提供重要的见解。这个缺失突变体的表型将从细胞活力、胚胎和胚胎后发育方面进行分析。如果零等位基因产生早期致命的表型，它将被携带在含有野生型基因染色体外阵列的品系中，这将允许通过分割阵列来产生纯合缺失动物。这种理解线虫发育过程中NFI功能的“反向遗传”方法还将允许测试脊椎动物的NFI基因是否可以补充线虫的基因，从而评估线虫突变在分析与脊椎动物发育有关的其他多基因家族中的作用。***

9612367 Gronostajski Nuclear Factor I (NFI) is one of a class of phylogenetically-conserved site-specific DNA-binding proteins that play an essential role in fundamental cellular processes including organismal development. These studies examine a simple developmental system in a organism containing a single NFI gene. The function of the NFI in the growth and development of C. elegans will be studied by the generation of deletion mutants in the NFI gene using Tcl targeted mutagenesis. The phenotype of homozygous null-mutants, and of organisms expressing specific mutated forms of NFI will yield important insights into how this family of evolutionarily-conserved DNA-binding proteins function in C. elegans and other organisms. The phenotype of this deletion mutant will be analyzed in respect to cell viability, embryonic, and post-embryonic development. If the null-allele generates an early lethal phenotype, it will be carried in a strain containing an extrachromosomal array of the wild-type gene which will allow the generation of homozygous deletion animals by partitioning of the array. This "reverse genetic" approach to understanding NFI function during C. elegans development will also allow testing whether the vertebrate NFI genes can complement the C. elegans gene and thus assess how useful C. elegans mutants may be in the analysis of other multigene families involved in vertebrate development. ***