Thomas K A, Schechter A N
Laboratory of Chemical Biology, National Institute of Arthritis, Metabolism, and Digestive Diseases, National Institutes of Health, Bethesda, Maryland 20014, USA.
Anal Biochem. 1978 Nov;91(1):209-23. doi: 10.1016/0003-2697(78)90833-3.
The cation of the salt ethidium bromide (3,8-diamino-5-ethyl-6-phenylphenanthridinium bromide) has been covalently linked to an agarose matrix through an intermediate 3,3'-diaminodipropylaminosuccinyl spacer arm. Partition binding and visible absorption spectral measurements on the gel were used to monitor the binding of transfer RNA to the covalently bound ethidium group. Direct fluorescence measurements of the formation of the gel-bound complex indicate that this binding involves the intercalation of the ethidium groups into the tRNA molecule. Dissociation of the ethidium-tRNA complex was monitored as a function of sodium chloride concentration by both direct solution spectral measurement of the released tRNA and by fluorescence quenching measurements of the dissociation of the intercalation complex. The derivatized gel has been shown to be capable of the fractionation of tRNA species by elution with a positive salt gradient under column flow conditions.
盐溴化乙锭（3,8 - 二氨基 - 5 - 乙基 - 6 - 苯基菲啶溴化物）的阳离子已通过中间的3,3'-二氨基二丙基氨基琥珀酰间隔臂与琼脂糖基质共价连接。对凝胶进行分配结合和可见吸收光谱测量，以监测转运RNA与共价结合的溴化乙锭基团的结合。对凝胶结合复合物形成的直接荧光测量表明，这种结合涉及溴化乙锭基团插入到tRNA分子中。通过对释放的tRNA进行直接溶液光谱测量以及对插入复合物解离进行荧光猝灭测量，监测溴化乙锭 - tRNA复合物的解离与氯化钠浓度的关系。已证明衍生化凝胶能够在柱流条件下通过用正盐梯度洗脱来分离tRNA种类。
