使用磷 - 32和磷 - 33同位素研究S1核酸酶对RNA - DNA杂交体的特异性。
The specificity of S1 nuclease toward RNA-DNA hybrids as studied using isotopes of phosphorus-32 and phosphorus-33.
作者信息
Wittelsberger S C, Hansen J N
出版信息
Nucleic Acids Res. 1977 Jun;4(6):1829-35. doi: 10.1093/nar/4.6.1829.
Abstract
Hybrids were formed from Bacillus cereus DNA and ribosomal RNA. They were treated with various combination of S1 nuclease and ribonuclease, and the molar ratios of the RNA and DNA moieties remaining in the treated hybrids were determined using a 32P-33P dual-label technique. It was found that both S1 nuclease and ribonuclease are required to give hybrid with RNA and DNA in a perfect 1:1 molar ratio. It was noted that the dual-label technique which employs orthophosphate as the sole phosphorus source for both labels gives unambiguous molar ratios and obviates the need to calculate specific activities, make quench corrections, or correct for base content.
摘要
杂种是由蜡样芽孢杆菌的DNA和核糖体RNA形成的。它们用S1核酸酶和核糖核酸酶的各种组合进行处理，并使用32P-33P双标记技术测定处理后的杂种中剩余的RNA和DNA部分的摩尔比。发现需要S1核酸酶和核糖核酸酶才能产生RNA和DNA摩尔比完美为1:1的杂种。注意到采用正磷酸盐作为两种标记的唯一磷源的双标记技术给出了明确的摩尔比，并且无需计算比活性、进行淬灭校正或校正碱基含量。
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