Wahl M C, Ramakrishnan B, Ban C, Chen X, Sundaralingam M
The Ohio State University, Laboratory of Biological Macromolecular Structure, Department of Biochemistry, Columbus 43210-1002, USA.
Acta Crystallogr D Biol Crystallogr. 1996 Jul 1;52(Pt 4):668-75. doi: 10.1107/S0907444996002788.
Protocols for the routine chemical synthesis and purification of milligram quantities of RNA and DNA-RNA chimeras meeting the demands of X-ray crystallography are described. An efficient screening protocol to test the crystallizability of the molecules and the optimization of the crystallization conditions are presented, so as to allow reproduction by others. Essentially the same crystallization conditions as for DNA oligomers can be employed for RNA crystallization. Specific examples involving alternating octamers, G/C-rich decamers, sequences with overhangs, and drug complexes of chimeras are discussed. Success of the methods is attested by the crystals obtained which diffract to high resolution.
本文描述了满足X射线晶体学要求的毫克级RNA和DNA-RNA嵌合体的常规化学合成及纯化方案。提出了一种有效的筛选方案,用于测试分子的结晶性并优化结晶条件,以便他人能够重复。RNA结晶基本上可以采用与DNA寡聚物相同的结晶条件。文中讨论了涉及交替八聚体、富含G/C的十聚体、有突出端的序列以及嵌合体药物复合物的具体实例。所获得的晶体能够衍射到高分辨率,证明了这些方法的成功。