SYNTHESIS OF BACKBONE DEUTERIUM LABELED [R(CGCGAA-UUCGCG)]2 AND HPLC PURIFICATION OF SYNTHETIC RNA

被引:12
作者
KHARE, D [1 ]
ORBAN, J [1 ]
机构
[1] UNIV MARYLAND,MARYLAND BIOTECHNOL INST,CTR ADV RES BIOTECHNOL,9600 GUDELSKY DR,ROCKVILLE,MD 20850
关键词
D O I
10.1093/nar/20.19.5131
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The chemical synthesis of backbone deuterium labelled [r(CGCGAAU*U*CGCG)]2 (U* = [5'-H-2]U) is described. An efficient purification procedure was developed using a polymeric reverse phase (PRP) HPLC column at 60-degrees-C. This procedure provided pure RNA dodecamer in the multi-milligram quantities (39% overall yield) necessary for dynamics studies using solid-state deuterium NMR. The purification method has been effectively applied to other RNA sequences and will assist biophysical studies which require relatively large quantities of RNA oligomers.
引用
收藏
页码:5131 / 5136
页数:6
相关论文
共 23 条
[1]   A SOLID-STATE DEUTERIUM NMR INVESTIGATION OF CONFORMATION AND ORDER IN MAGNETICALLY ORIENTED [D(CGCGAATTCGCG)]2 [J].
ALAM, TM ;
ORBAN, J ;
DROBNY, G .
BIOCHEMISTRY, 1990, 29 (41) :9610-9617
[2]   DEUTERIUM NMR INVESTIGATION OF BACKBONE DYNAMICS IN THE SYNTHETIC OLIGONUCLEOTIDE [D(CGCGAATTCGCG)]2 [J].
ALAM, TM ;
ORBAN, J ;
DROBNY, GP .
BIOCHEMISTRY, 1991, 30 (38) :9229-9237
[3]  
CARUTHERS MH, 1982, CHEM ENZYMATIC SYNTH, P71
[4]   SOLUTION STRUCTURE OF AN UNUSUALLY STABLE RNA HAIRPIN, 5'GGAC(UUCG)GUCC [J].
CHEONG, CJ ;
VARANI, G ;
TINOCO, I .
NATURE, 1990, 346 (6285) :680-682
[5]   SOLID-PHASE SYNTHESIS AND HIGH-RESOLUTION NMR-STUDIES OF 2 SYNTHETIC DOUBLE-HELICAL RNA DODECAMERS - R(CGCGAAUUCGCG) AND R(CGCGUAUACGCG) [J].
CHOU, SH ;
FLYNN, P ;
REID, B .
BIOCHEMISTRY, 1989, 28 (06) :2422-2435
[6]   HIGH-RESOLUTION NMR-STUDIES OF CHIMERIC DNA-RNA-DNA DUPLEXES, HETERONOMOUS BASE-PAIRING, AND CONTINUOUS BASE STACKING AT JUNCTIONS [J].
CHOU, SH ;
FLYNN, P ;
WANG, A ;
REID, B .
BIOCHEMISTRY, 1991, 30 (21) :5248-5257
[7]   MECHANISM OF INACTIVATION OF ESCHERICHIA-COLI AND LACTOBACILLUS-LEICHMANNII RIBONUCLEOTIDE REDUCTASES BY 2'-CHLORO-2'-DEOXYNUCLEOTIDES - EVIDENCE FOR GENERATION OF 2-METHYLENE-3(2H)-FURANONE [J].
HARRIS, G ;
ATOR, M ;
STUBBE, J .
BIOCHEMISTRY, 1984, 23 (22) :5214-5225
[8]   STRUCTURAL FEATURES THAT GIVE RISE TO THE UNUSUAL STABILITY OF RNA HAIRPINS CONTAINING GNRA LOOPS [J].
HEUS, HA ;
PARDI, A .
SCIENCE, 1991, 253 (5016) :191-194
[9]  
HNIKONOWICZ EP, 1992, J AM CHEM SOC, V114, P1082
[10]  
HNIKONOWICZ EP, 1992, NATURE, V355, P184