THE USE OF OLIGONUCLEOTIDE PROBES CONTAINING 2'-DEOXY-2'-FLUORONUCLEOSIDES FOR REGIOSPECIFIC CLEAVAGE OF RNA BY RNASE-H FROM ESCHERICHIA-COLI

被引：18

作者：

SCHMIDT, S

NIEMANN, A

KRYNETSKAYA, NF

ORETSKAYA, TS

METELEV, VG

SUCHOMLINOV, VV

SHABAROVA, ZA

CECH, D

机构：

[1] MV LOMONOSOV STATE UNIV,DEPT CHEM,MOSCOW 117234,USSR

[2] AN BELOZERSKII MOLEC BIOL & BIOORGAN CHEM LAB,MOSCOW,USSR

来源：

BIOCHIMICA ET BIOPHYSICA ACTA | 1992年 / 1130卷 / 01期

关键词：

MODIFIED OLIGONUCLEOTIDE; 2'-DEOXY-2'-FLUORONUCLEOSIDE; RNA-CLEAVAGE; DNA/RNA-CONFORMATION; RNASE-H;

D O I：

10.1016/0167-4781(92)90459-D

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：

Protected 2'-deoxy-2'-fluorouridine and 2'-deoxy-2'-fluorocytidine suitable for incorporation into oligonucleotides via the phosphoramidite approach have been prepared. Five modified and two unmodified oligonucleotides have been synthesized to investigate the regiospecific cleavage of a 5S RNA from Escherichia coli by RNase H. In order to show whether the modified oligonucleotides are able to hybridize with the RNA the physico-chemical properties (melting curves, CD spectra) of analogous DNA/oligodeoxyribonucleotide duplexes have been examined. The modified oligonucleotides are shown to form stable duplexes with a DNA-matrix which exist in an A-like form. Two of the modified probes containing four 2'-deoxy-2'-fluorocytidines or two 2'-deoxy-2'-fluorouridines direct the splitting by RNase H of only one phosphodiester bond of the RNA.

引用

页码：41 / 46

页数：6

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