DETERMINATION OF 7-METHYLGUANOSINE-CONTAINING 5'-TERMINI OF MESSENGER RIBONUCLEIC-ACID BY NAB[H-3]4 LABELING AND HIGH-PERFORMANCE LIQUID ANION-EXCHANGE CHROMATOGRAPHY

被引：16

作者：

BRETER, HJ ^{[1
]}

MALEK, LT ^{[1
]}

HELLMANN, GM ^{[1
]}

RHOADS, RE ^{[1
]}

机构：

[1] UNIV KENTUCKY,COLL MED,DEPT BIOCHEM,LEXINGTON,KY 40536

来源：

ANALYTICAL BIOCHEMISTRY | 1979年 / 98卷 / 01期

关键词：

D O I：

10.1016/0003-2697(79)90712-7

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A method is described for the determination of 5′-terminal methylated (cap) structures in unlabeled mRNA based on oxidation with NaIO4, reduction with NaB[3H]4, cleavage with P1 nuclease, and separation on a strong anion-exchange resin by high-performance liquid chromatography (hplc). Model compounds (cap 1 dinucleotides) were used to show that no structural alteration other than cleavage of the ribose ring of 7-methylguanosine occurred under the conditions used for oxidation and reduction. It was shown that the enzyme tobacco acid pyrophosphatase could be used to cleave cap dinucleotides containing unmodified or ring-opened ribose moieties and could also be used to release [3H]pm7G′ from NaB[3H]4-labeled rabbit globin mRNA. All five known cap 1 dinucleotides were resolved by hplc. The cap of rabbit globin mRNA was identified as m7Gpppm6Am, in agreement with other methods of determination. © 1979.

引用

页码：102 / 111

页数：10

共 32 条

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