用T4 RNA连接酶等摩尔添加寡核糖核苷酸。
Equimolar addition of oligoribonucleotides with T4 RNA ligase.
作者信息
Uhlenbeck O C, Cameron V
出版信息
Nucleic Acids Res. 1977 Jan;4(1):85-98. doi: 10.1093/nar/4.1.85.
Abstract
T4 induced RNA ligase will join equimolar concentrations of two oligoribonucleotides, (Ap)3C and p(Up) 5, to form a single product, (Ap)3Cp(Up) 5, in high yield. The presence of the 3' phosphate on p(Up)5 prevents the oligomer from adding to itself. The pH optimum of the reaction is about 7.5, but less of the undesirable adenylated intermediate, App(Up) 5, forms at pH 8.2. The reaction rate is a linear function of oligomer concentration from 3 micronM to 0.6 mM. The data suggest that T4 RNA ligase will be a useful enzyme for the synthesis of oligomers of defined sequence.
摘要
T4诱导的RNA连接酶能以高产率将等摩尔浓度的两种寡核糖核苷酸，即(Ap)3C和p(Up)5连接起来，形成单一产物(Ap)3Cp(Up)5。p(Up)5上3'磷酸基团的存在可防止该寡聚物自身连接。该反应的最适pH约为7.5，但在pH 8.2时，形成的不良腺苷酸化中间体App(Up)5较少。反应速率在寡聚物浓度从3微摩尔/升至0.6毫摩尔/升范围内是线性函数。这些数据表明，T4 RNA连接酶将是合成特定序列寡聚物的一种有用酶。
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