Digweed M, Pieler T, Kluwe D, Schuster L, Walker R, Erdmann V A
Eur J Biochem. 1986 Jan 2;154(1):31-9. doi: 10.1111/j.1432-1033.1986.tb09355.x.
An improved method for the isolation of a double-strand-specific RNase from snake venom is presented. This RNase, called CSV, was used to cleave yeast tRNAPhe and tRNA2Glu and tRNAfMet from Escherichia coli. In addition these RNAs and E. coli tRNAPhe were examined with the single-strand-specific nuclease S1. The results are discussed in terms of the specificity of CSV RNase and the structure of tRNAs. S1 nuclease digestions at increasing temperatures allowed the melting of tertiary and secondary structure to be monitored. 5S rRNA from E. coli, Thermoplasma acidophilum and the chloroplasts of Spinacia oleracea were digested with CSV and S1. The information these results give on the secondary-structural differences between different classes of 5S rRNA are discussed. Supporting evidence is found for tertiary interactions between hairpin loop c and internal loop d of eubacterial 5S rRNA.
本文介绍了一种从蛇毒中分离双链特异性核糖核酸酶的改进方法。这种核糖核酸酶称为CSV，用于切割酵母苯丙氨酸转运核糖核酸（tRNAPhe）、大肠杆菌的谷氨酰胺转运核糖核酸（tRNA2Glu）和甲硫氨酸起始转运核糖核酸（tRNAfMet）。此外，还使用单链特异性核酸酶S1对这些核糖核酸以及大肠杆菌的苯丙氨酸转运核糖核酸进行了检测。根据CSV核糖核酸酶的特异性和转运核糖核酸的结构对结果进行了讨论。在不断升高的温度下用S1核酸酶进行消化，可以监测三级和二级结构的解链情况。用CSV和S1对大肠杆菌、嗜热栖热菌和菠菜叶绿体的5S核糖体核糖核酸进行了消化。讨论了这些结果所提供的关于不同类别的5S核糖体核糖核酸二级结构差异的信息。发现了支持真细菌5S核糖体核糖核酸发夹环c和内环d之间三级相互作用的证据。
