铽作为RNA的固态探针

Gene Yonuschot, Donald Helman , George Mushrush , George Vande Woude, Gerry Robey
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引用次数: 11

摘要

本文继续了以往在固体状态下核酸-铽配合物分析方面的工作。报道了rna -铽(III)配合物的荧光激发和发射光谱。报道了RNA-terbium(III)和DNA-terbium(III)配合物在微孔过滤器上的荧光激发和发射光谱。与铽结合的DNA 100%被捕获在微孔过滤器上。采用sds萃取法从微孔过滤器捕获的dna -铽(III)配合物中回收脱氧核糖核酸。能量转移发生在从核酸中的碱基到铽离子,而实际的铽与核酸的结合是由于磷酸基团。同源多核糖核苷酸-铽络合物的相对荧光表明,鸟嘌呤部分负责大部分观察到的荧光。结合研究表明放射性铽对所有的均多核糖核苷酸具有相同的亲和力。利用固体DNA和RNA铽配合物的荧光测定DNA或RNA的皮摩尔量。
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Terbium as a solid-state probe for RNA

This paper continues previous work on the analysis of nucleic acid-terbium complexes in the solid state. The fluorescence excitation and emission spectra of the RNA-terbium(III) complex is reported. The fluorescence excitation and emission spectra of both the RNA-terbium(III) and DNA-terbium(III) complexes as trapped on millipore filters is reported. One hundred percent of the DNA combined with terbium was trapped on millipore filters. Deoxyribonucleic acid was recovered from DNA-terbium(III) complexes trapped on millipore filters using SDS-extraction.

Energy transfer was shown to occur from the bases in nucleic acids to the terbium ion, whereas the actual binding of terbium to nucleic acids was due to phosphate groups. The relative fluorescence of homopolyribonucleotide-terbium complexes showed that the guanine moiety was responsible for most of the observed fluorescence. Binding studies showed an equal affinity of radioactive terbium for all the homopolyribonucleotides. The fluorescence of solid-state DNA and RNA terbium complexes was used to measure picomole quantities of DNA or RNA.

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