枯草芽孢杆菌中tRNAMet-f、tRNAPhe和tRNATyr2的纯化与特性分析
Purification and characterization of tRNAMet-f, tRNAPhe and tRNATyr2 from Baccillus subtilis.
作者信息
Raettig R, Schmidt W, Mahal G, Kersten H, Arnold H H
出版信息
Biochim Biophys Acta. 1976 Jun 18;435(2):109-18. doi: 10.1016/0005-2787(76)90241-0.
Three tRNAs specific for methionine, phenylalanine and tyrosine were isolated from the total tRNA of Bacillus subtilis by chromatographic procedures using BD-cellulose and reversed-phase (5) chromatography. The acceptor activities of the purified tRNAs are 1160, 1260 and 1320 pmoles per A260nm unit for tRNAMetf, tRNAPhe and tRNATyr2 respectively. In tRNAMetf and tRNAPhe ribothymidine, pseudouridine and dihydrouridine are present, in addition, in tRNAPhe 7-methyguanosine and a 2'-O-methylated nucleoside were found. The modified nucleosides of tRNATyr2 are ribothymidine, pseudouridine, dihydrouridine, 4-thiouridine and 1-methyladenosine. The results suggest the presence of 2-methylthio-N6(delta 2-isopentenyl)adenosine in tRNAPhe and tRNATyr2. The thermal denaturation profiles of the three tRAN species are presented.
通过使用BD-纤维素和反相（5）色谱的色谱方法，从枯草芽孢杆菌的总tRNA中分离出了三种分别对甲硫氨酸、苯丙氨酸和酪氨酸特异的tRNA。纯化后的tRNA的接受活性分别为：tRNAMetf每A260nm单位1160皮摩尔，tRNAPhe每A260nm单位1260皮摩尔，tRNATyr2每A260nm单位1320皮摩尔。在tRNAMetf和tRNAPhe中存在核糖胸腺嘧啶、假尿嘧啶和二氢尿嘧啶，此外，在tRNAPhe中还发现了7-甲基鸟苷和一种2'-O-甲基化核苷。tRNATyr2的修饰核苷有核糖胸腺嘧啶、假尿嘧啶、二氢尿嘧啶、4-硫尿嘧啶和1-甲基腺苷。结果表明在tRNAPhe和tRNATyr2中存在2-甲硫基-N6（δ2-异戊烯基）腺苷。给出了这三种tRNA的热变性图谱。
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