Jett S D, Bear D G
Department of Cell Biology, University of New Mexico School of Medicine, Albuquerque 87131.
Proc Natl Acad Sci U S A. 1994 Jul 19;91(15):6870-4. doi: 10.1073/pnas.91.15.6870.
We present a technique, "snapshot blotting," for the electrophoretic transfer of nucleic acids and nucleoprotein complexes in gel electrophoresis bands onto highly stable carbon film-coated grids for imaging by electron microscopy. The method permits structural analysis of macromolecular species that have been resolved by a gel mobility-shift assay. To demonstrate the efficiency and integrity of the transfer process for a multiprotein-DNA assembly, we have imaged various species of a prokaryotic transcription complex, using the cleavage-defective EcoRI(Q111) protein as an orientation marker and as a blockade of transcription elongation. Snapshot blotting should be of great utility in the structural characterization of nucleic acids and protein-nucleic acid interactions.
我们介绍了一种名为“快照印迹法”的技术，用于在凝胶电泳条带中将核酸和核蛋白复合物电泳转移到用于电子显微镜成像的高度稳定的碳膜包被网格上。该方法允许对通过凝胶迁移率变动分析分离的大分子种类进行结构分析。为了证明多蛋白-DNA组装体转移过程的效率和完整性，我们使用切割缺陷型EcoRI（Q111）蛋白作为方向标记和转录延伸的阻断剂，对原核转录复合物的各种种类进行了成像。快照印迹法在核酸和蛋白质-核酸相互作用的结构表征中应具有很大的实用性。
