一种改进的用于分离和定量转移核糖核酸修饰核苷的方法。
An improved method for the separation and quantitation of the modified nucleosides of transfer RNA.
作者信息
Rogg H, Brambilla R, Keith G, Staehelin M
出版信息
Nucleic Acids Res. 1976 Jan;3(1):285-95. doi: 10.1093/nar/3.1.285.
Abstract
A method is described which allows a very efficient determination of the modified nucleosides of tRNA. The technique involves enzymatic degradation of the tRNA to nucleosides at pH 7.6 and their separation by two-dimensional thin-layer chromatography on cellulose-coated aluminum foils. Based on the analysis of two mammalian tRNAs it is shown that the technique is suitable for the determination of chemically unstable nucleosides as well as the ribose-methylated compounds. At least 36 of the 45 known modified nucleosides can be separated and quantitatively determined by the method described. This procedure is especially suitable for the estimation of the nucleoside composition of unlabeled tRNAs as well as for studying the post-transcriptional modifications of tRNA.
摘要
本文描述了一种能非常高效地测定tRNA修饰核苷的方法。该技术包括在pH 7.6条件下将tRNA酶解为核苷，并通过在涂有纤维素的铝箔上进行二维薄层色谱法对其进行分离。基于对两种哺乳动物tRNA的分析表明，该技术适用于测定化学不稳定核苷以及核糖甲基化化合物。通过所述方法可以分离并定量测定已知的45种修饰核苷中的至少36种。该程序特别适用于估计未标记tRNA的核苷组成以及研究tRNA的转录后修饰。
相似文献
1
An improved method for the separation and quantitation of the modified nucleosides of transfer RNA.一种改进的用于分离和定量转移核糖核酸修饰核苷的方法。
Nucleic Acids Res. 1976 Jan;3(1):285-95. doi: 10.1093/nar/3.1.285.
2
Ribonucleoside analysis by reversed-phase high-performance liquid chromatography.反相高效液相色谱法进行核糖核苷分析
J Chromatogr. 1989 Jun 2;471:3-36. doi: 10.1016/s0021-9673(00)94152-9.
3
Improved separation of modified nucleosides from tRNA hydrolysates: the patterns of tRNA methylation in rat tissues.
J Biochem Biophys Methods. 1979 Oct;1(5):287-98. doi: 10.1016/0165-022x(79)90003-4.
4
A rapid assay technique for RNA ribose methylases.一种用于RNA核糖甲基化酶的快速检测技术。
Nucleic Acids Res. 1975 Apr;2(4):603-11. doi: 10.1093/nar/2.4.603.
5
6
Determination of nucleoside composition of ribonucleic acid by thin-layer chromatography.
Anal Biochem. 1968 Oct 24;25(1):55-60. doi: 10.1016/0003-2697(68)90080-8.
7
Separation of the methylated nucleoside consituents of ribonucleic acid by two-dimensional thin-layer chromatography.
J Chromatogr. 1975 Sep 3;111(2):403-8. doi: 10.1016/s0021-9673(00)99290-2.
8
High resolution chromatography of ribonucleosides and its application to RNA analysis.核糖核苷的高分辨率色谱法及其在RNA分析中的应用。
Biomed Chromatogr. 1989 Nov;3(6):246-50. doi: 10.1002/bmc.1130030604.
9
Separation of methylated nucleosides by high-pressure liquid chromatography. The pattern of tRNA methylation in stimulated rat lymphocytes.
Anal Biochem. 1976 Nov;76(l):342-50. doi: 10.1016/0003-2697(76)90292-x.
10
Quantitative enzymatic hydrolysis of tRNAs: reversed-phase high-performance liquid chromatography of tRNA nucleosides.tRNA的定量酶促水解：tRNA核苷的反相高效液相色谱法
J Chromatogr. 1982 Jul 9;230(2):297-308.
引用本文的文献
1
The occurrence order and cross-talk of different tRNA modifications.不同 tRNA 修饰的发生顺序和串扰。
Sci China Life Sci. 2021 Sep;64(9):1423-1436. doi: 10.1007/s11427-020-1906-4. Epub 2021 Apr 19.
2
The methyl group of the N6-methyl-N6-threonylcarbamoyladenosine in tRNA of Escherichia coli modestly improves the efficiency of the tRNA.大肠杆菌转运核糖核酸（tRNA）中N6-甲基-N6-苏氨酰氨甲酰腺苷的甲基基团适度提高了tRNA的效率。
J Bacteriol. 1998 Apr;180(7):1808-13. doi: 10.1128/JB.180.7.1808-1813.1998.
3
A novel method for the determination of post-transcriptional modification in RNA by mass spectrometry.一种通过质谱法测定RNA转录后修饰的新方法。
Nucleic Acids Res. 1993 Sep 25;21(19):4577-85. doi: 10.1093/nar/21.19.4577.
4
Nucleotide sequences of Acanthamoeba castellanii 5S and 5.8S ribosomal ribonucleic acids: phylogenetic and comparative structural analyses.卡氏棘阿米巴5S和5.8S核糖体核糖核酸的核苷酸序列：系统发育和比较结构分析
Nucleic Acids Res. 1981 Jul 24;9(14):3321-34. doi: 10.1093/nar/9.14.3321.
5
Nucleotide sequence of an exceptionally long 5.8S ribosomal RNA from Crithidia fasciculata.来自纤细短膜虫的超长5.8S核糖体RNA的核苷酸序列。
Nucleic Acids Res. 1982 Mar 25;10(6):2085-92. doi: 10.1093/nar/10.6.2085.
6
The transfer RNA of certain Enterobacteriacae contain 2-methylthiozeatin riboside (ms2io6A) an isopentenyl adenosine derivative.某些肠杆菌科细菌的转运核糖核酸含有2-甲硫基玉米素核苷（ms2io6A），一种异戊烯基腺苷衍生物。
Nucleic Acids Res. 1982 Sep 25;10(18):5663-72. doi: 10.1093/nar/10.18.5663.
7
Nucleotide sequence of valine tRNA mo5UAC from bacillus subtilis.来自枯草芽孢杆菌的缬氨酸tRNA mo5UAC的核苷酸序列。
Nucleic Acids Res. 1982 Jan 22;10(2):715-8. doi: 10.1093/nar/10.2.715.
8
Purification and properties of several transfer RNA methyltransferases from S. typhimurium.鼠伤寒沙门氏菌中几种转移RNA甲基转移酶的纯化及特性
Mol Cell Biochem. 1981 Apr 27;36(2):95-104. doi: 10.1007/BF02354908.
9
Novel E. coli mutants deficient in biosynthesis of 5-methylaminomethyl-2-thiouridine.缺乏5-甲基氨甲基-2-硫代尿苷生物合成的新型大肠杆菌突变体。
Nucleic Acids Res. 1984 Apr 25;12(8):3521-34. doi: 10.1093/nar/12.8.3521.
10
Undermodification in the first position of the anticodon of supG-tRNA reduces translational efficiency.supG - tRNA反密码子第一位的修饰不足会降低翻译效率。
Mol Gen Genet. 1984;196(2):194-200. doi: 10.1007/BF00328050.
本文引用的文献
1
Studies on polynucleotides, lxviii the primary structure of yeast phenylalanine transfer RNA.多核苷酸研究，第六十八篇：酵母苯丙氨酸转移核糖核酸的一级结构
Proc Natl Acad Sci U S A. 1967 Mar;57(3):751-8. doi: 10.1073/pnas.57.3.751.
2
Chromatographic studies of nucleic acids. 4. The nucleic acid of the turnip yellow mosaic virus including a note on the nucleic acid of the tomato bushy stunt virus.核酸的色谱研究。4. 芜菁黄花叶病毒的核酸，包括关于番茄丛生矮化病毒核酸的一则注释。
Biochem J. 1951 Sep;49(4):401-6. doi: 10.1042/bj0490401.
3
The separation and estimation of ribonucleotides in minute quantities.微量核糖核苷酸的分离与测定
J Biol Chem. 1950 Sep;186(1):37-50.
4
THIN-LAYER CHROMATOGRAPHY OF NUCLEOTIDES, NUCLEOSIDES AND NUCLEIC ACID BASES.核苷酸、核苷及核酸碱基的薄层色谱法
Biochim Biophys Acta. 1965 Jan 11;95:1-7. doi: 10.1016/0005-2787(65)90204-2.
5
STRUCTURE OF A RIBONUCLEIC ACID.核糖核酸的结构
Science. 1965 Mar 19;147(3664):1462-5. doi: 10.1126/science.147.3664.1462.
6
ON THE SPECIFICITY OF RNAASE T1.
Biochim Biophys Acta. 1964 Jul 22;87:493-5. doi: 10.1016/0926-6550(64)90121-5.
7
SOME NUCLEOTIDE SEQUENCES FROM PARTIALLY PURIFIED TRANSFER RIBONUCLEIC ACIDS.部分纯化的转移核糖核酸的一些核苷酸序列
Biochim Biophys Acta. 1964 Jun 22;87:199-211. doi: 10.1016/0926-6550(64)90216-6.
8
The sequence of phenylalanine tRNA from E. coli.来自大肠杆菌的苯丙氨酸转运核糖核酸的序列。
FEBS Lett. 1969 Jun;3(4):275-278. doi: 10.1016/0014-5793(69)80157-2.
9
Primary sequence of glutamic acid tRNA II from Escherichia coli.来自大肠杆菌的谷氨酸tRNA II的一级序列。
FEBS Lett. 1972 Feb 1;20(2):239-241. doi: 10.1016/0014-5793(72)80804-4.
10
Analysis of methylated constituents from RNA by thin-layer chromatography.通过薄层色谱法分析RNA中的甲基化成分。
Biochim Biophys Acta. 1967 Apr 18;138(2):430-2. doi: 10.1016/0005-2787(67)90504-7.
Zotero 插件