Chan Rene C, Forbes Douglass I
Division of Biological Sciences, University of California, San Diego, USA.
Methods Mol Biol. 2006;322:289-300. doi: 10.1007/978-1-59745-000-3_20.
Nuclear import is a critical process for the cell: molecules are selectively permitted into the nuclear interior where the sheltered genome resides. The process of nuclear import can be biochemically studied in vitro using nuclei reconstituted from Xenopus egg extract components and Xenopus sperm chromatin. This in vitro system allows for the visualization of nuclear import by monitoring the accumulation of fluorescent nuclear import substrates in the reconstituted nuclei. A powerful aspect of the system is that "biochemically mutant" nuclei can be readily generated, either by immunodepletion of proteins from or addition of proteins to the reaction. This ability allows ascertainment of the role of specific proteins in nuclear import.
分子被选择性地允许进入含有受保护基因组的细胞核内部。核输入过程可以在体外通过使用从非洲爪蟾卵提取物成分和非洲爪蟾精子染色质重构的细胞核进行生物化学研究。这个体外系统通过监测荧光核输入底物在重构细胞核中的积累来实现核输入的可视化。该系统的一个强大之处在于，可以通过从反应中免疫去除蛋白质或向反应中添加蛋白质，轻松生成“生物化学突变”的细胞核。这种能力使得确定特定蛋白质在核输入中的作用成为可能。
