SGER: Membrane-Elution and Differential Display Analysis of Cyclic Gene Expression

SGER：循环基因表达的膜洗脱和差异显示分析

基本信息

批准号：
0323346
负责人：
Stephen Cooper
金额：
$ 5.5万
依托单位：
Regents of the University of Michigan - Ann Arbor
依托单位国家：
美国
项目类别：
Standard Grant
财政年份：
2004
资助国家：
美国
起止时间：
2004-05-01 至 2006-09-30
项目状态：
已结题

来源：
https://www.nsf.gov/awardsearch/showAward?AWD_ID=0323346&HistoricalAwards=false
关键词：
SGER Membrane Elution Differential Display

项目摘要

The experimental work in the project will measure gene expression during the division cycle of eukaryotic, mammalian cells of both human and mouse origin. The newly developed membrane elution (or baby machine) method for producing undisturbed, synchronous cultures of eukaryotic cells will be used to analyze the precise pattern and timing of expression of different genes during the mammalian division cycle. This method produces unperturbed synchronized cells that exhibit three generations of synchronized divisions with the proper DNA contents and cell sizes observed throughout the successive cell cycles. The hypothesis to be tested is that there are few cell-cycle-regulated genes in eukaryotic cells, and in particular, there are no G1-phase specific patterns of gene expression. In the membrane-elution method, exponentially growing, unperturbed cells are bound to a membrane. The membrane is washed with warm medium so cells grow and divide on the membrane. Only newborn cells are released from the membrane by division. A short collection period yields a synchronized culture. The broader impact of this work is that the entire field of cell biology, arguably depends on a correct understanding of the nature of the cell cycle. If the hypothesis proves correct, it could have a major impact on how we understand the cell cycle.

该项目的实验工作将测量人类和小鼠来源的真核哺乳动物细胞分裂周期中的基因表达。新开发的膜洗脱（或婴儿机器）方法用于产生不受干扰的同步真核细胞培养物，将用于分析哺乳动物分裂周期中不同基因表达的精确模式和时间。该方法产生不受干扰的同步细胞，这些细胞表现出三代同步分裂，并在整个连续细胞周期中观察到适当的 DNA 含量和细胞大小。待检验的假设是真核细胞中细胞周期调节基因很少，特别是不存在 G1 期特异性基因表达模式。在膜洗脱方法中，指数生长的、未受干扰的细胞与膜结合。用温热的培养基清洗膜，使细胞在膜上生长和分裂。只有新生细胞通过分裂从膜中释放出来。较短的收集期可产生同步培养。这项工作更广泛的影响是，整个细胞生物学领域可以说取决于对细胞周期本质的正确理解。如果这个假设被证明是正确的，它可能会对我们如何理解细胞周期产生重大影响。