Boguski M S, Hieter P A, Levy C C
J Biol Chem. 1980 Mar 10;255(5):2160-3.
Rapid RNA sequencing technology was used to determine if the base specificities of an RNase recently purified from chicken liver would prove useful for RNA sequence analysis. Escherichia coli 5 S [5'-32P]rRNA or yeast 5.8 S [5'-32P]rRNA was digested with the enzyme and this digest, along with digests derived from RNases of known specificity (U2, T1, T2) were subjected to electrophoresis through denaturing polyacrylamide slab gels. Following autoradiography, the banding patterns arising from the activity of each enzyme were compared, and the base specificity of the unknown RNase was established. The chicken liver RNase was found to have a marked preference for phosphodiester bonds containing cytidylic acid residues, a property which should make the enzyme useful for distinguishing between pyrimidines in RNA sequencing.
利用快速RNA测序技术来确定最近从鸡肝中纯化出的一种核糖核酸酶的碱基特异性是否对RNA序列分析有用。用该酶消化大肠杆菌5S [5'-32P] rRNA或酵母5.8S [5'-32P] rRNA,并且将该消化产物以及源自已知特异性核糖核酸酶(U2、T1、T2)的消化产物通过变性聚丙烯酰胺平板凝胶进行电泳。放射自显影后,比较每种酶活性产生的条带模式,并确定未知核糖核酸酶的碱基特异性。发现鸡肝核糖核酸酶对含有胞苷酸残基的磷酸二酯键有明显偏好,这一特性应使该酶在RNA测序中区分嘧啶时有用。