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四环素阻遏物/操纵基因复合物的结晶及初步X射线分析

Crystallization and preliminary X-ray analysis of the Tet-repressor/operator complex.

作者信息

Orth P, Alings C, Schnappinger D, Saenger W, Hinrichs W

机构信息

Institut für Kristallographie, Freie Universität Berlin, Takustrasse 6, D-14195 Berlin, Germany.

出版信息

Acta Crystallogr D Biol Crystallogr. 1998 Jan 1;54(Pt 1):99-100. doi: 10.1107/s0907444997007646.

Abstract

Three crystal forms of the repressor protein TetR class D in complex with the palindromic 17 bp operator sequence containing T overhangs on both sides were obtained by hanging-drop vapor-diffusion methods using PEG 4000 and PEG monomethylether 5000 as precipitants. Although the crystallization conditions were very similar, up to three different crystal forms were observed in the same drop. The space groups are monoclinic C2, P21 and hexagonal P6122. The asymmetric units of the latter two crystal forms contain one repressor-operator complex. The crystal structures of these forms were solved by molecular replacement using the Tet-repressor molecule of the complex with tetracycline as a search model.

摘要

采用悬滴气相扩散法,以聚乙二醇4000(PEG 4000)和聚乙二醇单甲醚5000(PEG monomethylether 5000)作为沉淀剂,获得了阻遏蛋白D类与两侧带有T突出端的17bp回文操纵序列形成的三种晶体形式。尽管结晶条件非常相似,但在同一液滴中观察到了多达三种不同的晶体形式。空间群分别为单斜晶系C2、P21和六方晶系P6122。后两种晶体形式的不对称单元包含一个阻遏蛋白-操纵序列复合物。这些形式的晶体结构通过分子置换法解析,使用与四环素复合物的Tet阻遏蛋白分子作为搜索模型。

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