Dowty M E, Williams P, Zhang G, Hagstrom J E, Wolff J A
Department of Pediatrics, Waisman Center, University of Wisconsin, Madison 53705, USA.
Proc Natl Acad Sci U S A. 1995 May 9;92(10):4572-6. doi: 10.1073/pnas.92.10.4572.
These studies were initiated to elucidate the mechanism of DNA nuclear transport in mammalian cells. Biotin- or gold-labeled plasmid and plasmid DNA expression vectors for Escherichia coli beta-galactosidase or firefly luciferase were microinjected into the cytoplasm of primary rat myotubes in culture. Plasmid DNA was expressed in up to 70% of the injected myotubes, which indicates that it entered intact, postmitotic nuclei. The nuclear transport of plasmid DNA occurred through the nuclear pore by a process common to other large karyophilic macromolecules. The majority of the injected plasmid DNA was sequestered by cytoplasmic elements. This understanding of plasmid DNA nuclear transport provides a basis for increasing the efficiency of gene transfer.
开展这些研究是为了阐明哺乳动物细胞中DNA核转运的机制。将生物素或金标记的质粒以及用于大肠杆菌β-半乳糖苷酶或萤火虫荧光素酶的质粒DNA表达载体显微注射到培养的原代大鼠肌管的细胞质中。高达70%的注射肌管中表达了质粒DNA，这表明它完整地进入了有丝分裂后的细胞核。质粒DNA的核转运通过核孔，其过程与其他大型亲核大分子相同。大部分注射的质粒DNA被细胞质成分隔离。对质粒DNA核转运的这种理解为提高基因转移效率提供了基础。
