通过聚丙烯酰胺凝胶电泳测定核糖核酸的分子量。构象变化的影响。
The determination of the molecular weight of ribonucleic acid by polyacrylamide-gel electrophresis. The effects of changes in conformation.
作者信息
Loening U E
出版信息
Biochem J. 1969 Jun;113(1):131-8. doi: 10.1042/bj1130131.
Abstract
- The effects of changes in experimental conditions on the mobility of RNA in polyacrylamide-gel electrophoresis were investigated. 2. The linear relation between log(molecular weight) and electrophoretic mobility was shown to be independent within limits of salt or gel concentration. 3. The relative mobility of RNA with low content of guanylic acid and cytidylic acid residues was decreased in low-ionic-strength buffer. This was related to a small relative decrease in sedimentation coefficient. 4. However, Mg(2+) ion caused almost no increase in mobility although it was associated with large increases in sedimentation coefficient. This suggested opposing actions of Mg(2+) ion on the size and effective charge of the RNA. 5. It is concluded that the method provides a satisfactory measurement of molecular weight, which is almost independent of the nucleotide composition of RNA at moderate salt concentrations.
摘要
- 研究了实验条件的变化对RNA在聚丙烯酰胺凝胶电泳中迁移率的影响。2. 结果表明，在盐浓度或凝胶浓度的一定范围内，log（分子量）与电泳迁移率之间的线性关系是独立的。3. 在低离子强度缓冲液中，鸟苷酸和胞苷酸残基含量低的RNA的相对迁移率降低。这与沉降系数的相对小幅降低有关。4. 然而，Mg(2+)离子虽然与沉降系数的大幅增加有关，但几乎没有引起迁移率的增加。这表明Mg(2+)离子对RNA的大小和有效电荷有相反的作用。5. 得出的结论是，该方法能对分子量进行令人满意的测量，在中等盐浓度下，该测量几乎与RNA的核苷酸组成无关。
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