ANOMALOUS ELECTROPHORESIS OF DEOXYRIBONUCLEIC-ACID RESTRICTION FRAGMENTS ON POLYACRYLAMIDE GELS
ANOMALOUS ELECTROPHORESIS OF DEOXYRIBONUCLEIC-ACID RESTRICTION FRAGMENTS ON POLYACRYLAMIDE GELS
复制标题
DOI：
10.1021/bi00295a023
复制
发表时间：
1983-01-01
期刊：
影响因子：
2.9
通讯作者：
STELLWAGEN, NC
中科院分区：
文献类型：
--
作者：
STELLWAGEN, NC
本文对从pBR 322和p82 - 6B两个质粒中获得的DNA限制性片段进行了聚丙烯酰胺凝胶电泳。研究的变量是MW、凝胶浓度、温度和电场强度。较大片段（> 800个碱基对）的阻滞系数与分子量无关。最小碎片的延迟系数（≤ 0.000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000 300个碱基对）与分子量1/3和片段的平均几何半径成正比。所有片段的相对迁移率的对数也与MW 1/3成比例。某些片段在聚丙烯酰胺凝胶上的异常迁移可转移到由不同限制性内切酶产生的片段中。在较低的温度下，异常迁移增强，并在温度升高时消失。当二聚化时，错误迁移的片段缓慢迁移甚至更错误;二聚化正常迁移的片段导致二聚化片段的正常迁移。发现异常迁移的片段位于pBR 322环的不同区域。
A detailed study was made of the polyacrylamide gel electrophoresis of DNA restriction fragments obtained from 2 plasmids, pBR322 and p82-6B. Variables studied were MW, gel concentration, temperature and electric field strength. The retardation coefficients of the larger fragments (> 800 base pairs) were independent of MW. The retardation coefficients of the smallest fragments (.ltoreq. 300 base pairs) were proportional to MW1/3 and to the mean geometric radii of the fragments. The logarithm of the relative mobility of all fragments was also proportional to MW1/3. The anomalous migration of certain fragments on polyacrylamide gels is transportable into fragments generated by different restriction enzymes. Anomalous migration was enhanced at lower temperatures and disappeared upon increasing the temperature. A fragment which migrated anomalously slowly migrated even more anomalously when dimerized; dimerizing a normally migrating fragment resulted in the normal migration of the dimerized fragment. Anomalously migrating fragments were found to be localized in distinct regions of the pBR322 circle.