White B N, Dunn R, Gillam I, Tener G M, Armstrong D J, Skoog F, Frihart C R, Leonard N J
J Biol Chem. 1975 Jan 25;250(2):515-21.
Crude tRNA from adult Drosophila melanogaster was fractionated on bensoylated-diethylaminoethyl cellulose columns. The eluate was assayed for both amino acid acceptance and cytokinin activity. Most of the cytokinin activity was associated with a peak of serine acceptance. The five major serine tRNAs were purified by chromatography on benzoylated-dietyhlaminoethyl cellulose and reversed phase chromatography-5 columns. The major species, tRNA7-Ser was isolated from this tRNA and was shown to be N-6-(delta-2-isopentenyl)adenosine (i-6A) on the basis of ultraviolet and mass spectral data. The nucleoside somposition of all five serine tRNAs was determined directly and by the 3-H derivative method. They all contain pseudouridine, ribothymidine, 1-methyladenosine, 5-methylcytosine, N-2-dimethylguanosine, 5, 6-hydrouridine, and 3-methylcytosine, while two contain an unidentified nucleoside, and one containes 1-methylguanosine. These techniques also confirmed the presence of i-6A in tRNA7-Ser as well as showing its presence in tRNA6-Ser and tRNA4-Ser. These three tRNA-Ser species exhibit marked changes in elution from reversed phase chromatography-5 columns as a function of temperature and this may be related to their minor base composition. The tRNAs-Ser were bound to ribosomes in response to the following triplets: tRNA2-Ser, AGU, AGC; tRNA4-Ser, UCG; tRNA5-Ser, AGU, AGC; tRNA7-Ser, UCG.
从成年黑腹果蝇中提取的粗制tRNA在苯甲酰化二乙氨基乙基纤维素柱上进行分级分离。对洗脱液进行氨基酸接受能力和细胞分裂素活性检测。大部分细胞分裂素活性与丝氨酸接受峰相关。通过在苯甲酰化二乙氨基乙基纤维素柱上的色谱分离和反相色谱-5柱，纯化了5种主要的丝氨酸tRNA。从该tRNA中分离出主要种类tRNA7-Ser，并根据紫外和质谱数据表明其为N-6-（δ-2-异戊烯基）腺苷（i-6A）。直接并通过3-H衍生物法测定了所有5种丝氨酸tRNA的核苷组成。它们都含有假尿苷、核糖胸腺嘧啶、1-甲基腺苷、5-甲基胞嘧啶、N-2-二甲基鸟苷、5,6-二氢尿苷和3-甲基胞嘧啶，其中两种含有一种未鉴定的核苷，一种含有1-甲基鸟苷。这些技术还证实了i-6A在tRNA7-Ser中的存在，以及在tRNA6-Ser和tRNA4-Ser中的存在。这三种tRNA-Ser种类在反相色谱-5柱上的洗脱随温度有显著变化，这可能与其稀有碱基组成有关。丝氨酸tRNA响应以下三联体与核糖体结合：tRNA2-Ser，AGU，AGC；tRNA4-Ser，UCG；tRNA5-Ser，AGU，AGC；tRNA7-Ser，UCG。
