Curved DNA molecules migrate anomalously slowly in free solution.
Curved DNA molecules migrate anomalously slowly in free solution.
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DOI：
10.1093/nar/gki748
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发表时间：
2005
影响因子：
14.9
通讯作者：
Stellwagen NC
中科院分区：
文献类型：
--
作者：
Stellwagen E;Lu Y;Stellwagen NC
用毛细管电泳法测定了SV40微染色体VP1基因的弯曲DNA限制性片段在聚丙烯酰胺凝胶和无水溶液中的凝胶迁移率。该片段的表观弯曲角度为46±2°，位于SV40序列位置1922±2bp[Lu Y.J.，Weers B.D.和Stellwagen N.C.(2005)BiPhys]。J.，88,1191-1206]。表观弯曲中心周围的“曲率模块”包含五个间隔不均匀的A和T形束，它们是观测到的曲率的原因。在曲率模块中包含至少一个A-链的亲本199碱基片段和序列突变体在自由溶液中以及在聚丙烯酰胺凝胶中迁移异常缓慢。因此，观察到弯曲的DNA分子在聚丙烯酰胺凝胶中的异常缓慢的迁移率部分是由于它们在自由溶液中的异常缓慢的迁移率。对凝胶和游离溶液迁移率下降的分析表明，每个A-或T-链对199个碱基片段及其A-链突变体的曲率有独立的贡献，但不是相等的。每个A-或T-道对观测曲率的相对贡献取决于其相对于曲率模块中第一个A-道的间距。
The electrophoretic mobility of a curved DNA restriction fragment taken from the VP1 gene in the SV40 minichromosome has been measured in polyacrylamide gels and free solution, using capillary electrophoresis. The 199 bp restriction fragment has an apparent bend angle of 46 ± 2° located at SV40 sequence position 1922 ± 2 bp [Lu Y.J., Weers B.D. and Stellwagen N. C. (2005) Biophys. J., 88, 1191–1206]. The ‘curvature module’ surrounding the apparent bend center contains five unevenly spaced A- and T-tracts, which are responsible for the observed curvature. The parent 199 bp fragment and sequence mutants containing at least one A-tract in the curvature module migrate anomalously slowly in free solution, as well as in polyacrylamide gels. Hence, the anomalously slow mobilities observed for curved DNA molecules in polyacrylamide gels are due in part to their anomalously slow mobilities in free solution. Analysis of the gel and free solution mobility decrements indicates that each A- or T-tract contributes independently, but not equally, to the curvature of the 199 bp fragment and its A-tract mutants. The relative contribution of each A- or T-tract to the observed curvature depends on its spacing with respect to the first A-tract in the curvature module.
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