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

该项目的实验工作将测量人类和小鼠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）方法用于生产真核细胞的不受干扰的同步培养，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。这种方法产生不受干扰的同步细胞，表现出三代同步分裂，在整个连续的细胞周期中观察到适当的DNA含量和细胞大小。要验证的假设是真核细胞中很少有细胞周期调控基因，特别是没有g1期特异性的基因表达模式。在膜洗脱法中，指数生长的、未受干扰的细胞被结合到膜上。用温暖的培养基冲洗膜，使细胞在膜上生长和分裂。只有新生细胞通过分裂从细胞膜中释放出来。较短的收集周期产生同步区域性。这项工作的更广泛的影响是，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。

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.