論文 - 詳細
| RRC ID | 35202 |
|---|---|
| 著者 | Celesnik H, Deana A, Belasco JG. |
| タイトル | PABLO analysis of RNA: 5'-phosphorylation state and 5'-end mapping. |
| ジャーナル | Methods Enzymol |
| Abstract |
Recent studies have revealed that 5'-end-dependent RNA degradation in prokaryotes is triggered by pyrophosphate removal from the 5'-terminus to generate a monophosphorylated intermediate that is readily degraded. This chapter describes how to examine the 5'-phosphorylation state of any specific bacterial RNA by PABLO analysis. The method is based on the ability of monophosphorylated, but not triphosphorylated, RNA 5'-ends to undergo splinted ligation to a DNA oligonucleotide when juxtaposed by base pairing to a bridging oligonucleotide. PABLO analysis not only makes it possible to quantify the proportion of a particular RNA that is monophosphorylated in bacterial cells but also provides a more reliable method than primer extension for high-resolution mapping of RNA 5'-termini. |
| 巻・号 | 447 |
| ページ | 83-98 |
| 公開日 | 2008-1-1 |
| DOI | 10.1016/S0076-6879(08)02205-2 |
| PII | S0076-6879(08)02205-2 |
| PMID | 19161839 |
| MeSH | Base Sequence Blotting, Northern Diphosphates / isolation & purification Electrophoresis, Polyacrylamide Gel Molecular Sequence Data Phosphorylation RNA, Messenger / chemistry RNA, Messenger / metabolism* Reverse Transcriptase Polymerase Chain Reaction |
| IF | 1.394 |
| 引用数 | 23 |
| WOS 分野 | BIOCHEMICAL RESEARCH METHODS BIOCHEMISTRY & MOLECULAR BIOLOGY |
| リソース情報 | |
| 原核生物(大腸菌) | Review? |