Purification and Analysis of A/U-rich RNA Binding Protein Using Immobilized RNA by EDC Crosslinking
文献类型：期刊论文
| 作者 | Sun, DQ; Wang, Y; Liu, DG |
| 刊名 | PROGRESS IN BIOCHEMISTRY AND BIOPHYSICS
 |
| 出版日期 | 2013 |
| 卷号 | 40期号:4页码:386-391 |
| 关键词 | C/EBP beta 3 ' UTR A/U-rich RNA hnRNPC interaction between nucleic acid and protein |
| 通讯作者 | Liu, DG (reprint author), Chinese Acad Sci, State Key Lab Mol Biol, Inst Biochem & Cell Biol, Shanghai Inst Biol Sci, Shanghai 200031, Peoples R China.,dgliu@sibcb.ac.cn |
| 英文摘要 | An important way for RNA to implement its biological function in cells is the binding to proteins. Thus the analysis of RNA binding proteins in cells is necessary for the investigation of RNA's functions. So far, there have been various methods to identify RNA binding proteins. In this study, we describe a novel method, i.e. to immobilize an A/U-rich RNA to a solid medium amine M-270 magnetic beads by coupling them in a direct condensation reaction using the soluble carbodiimide reagent EDC, and then to isolate RNA binding proteins from cell lysate by RNA affinity chromatography using the immobilized RNA. The RNA specific binding proteins were then identified by SDS-PAGE, mass spectrometry and Western blotting, such as hnRNPC which bound specifically to the A/U-rich RNA of C/EBP beta 3'UTR. Colocalization in vivo between A/U-rich RNA and the identified protein hnRNPC was shown by fluorescence in situ hybridization using RNA molecular beacon and anti-hnRNPC antibody and confocal microscopy. Our results indicated that the direct immobilization of RNA with EDC is highly efficient, simple and easy. |
| 学科主题 | Biochemistry & Molecular Biology; Biophysics |
| 类目[WOS] | Biochemistry & Molecular Biology ; Biophysics |
| 关键词[WOS] | TUMOR SUPPRESSION ACTIVITY ; EXPRESSION ; MECHANISM ; SEQUENCES ; REVERSION ; REGION |
| 收录类别 | SCI |
| 语种 | 中文 |
| WOS记录号 | WOS:000318059500010 |
| 版本 | 出版稿 |
| 源URL | [http://202.127.25.143/handle/331003/377]  |
| 专题 | 上海生化细胞研究所_上海生科院生化细胞研究所 |
| 推荐引用方式 GB/T 7714 | Sun, DQ,Wang, Y,Liu, DG. Purification and Analysis of A/U-rich RNA Binding Protein Using Immobilized RNA by EDC Crosslinking[J]. PROGRESS IN BIOCHEMISTRY AND BIOPHYSICS,2013,40(4):386-391. |
| APA | Sun, DQ,Wang, Y,&Liu, DG.(2013).Purification and Analysis of A/U-rich RNA Binding Protein Using Immobilized RNA by EDC Crosslinking.PROGRESS IN BIOCHEMISTRY AND BIOPHYSICS,40(4),386-391. |
| MLA | Sun, DQ,et al."Purification and Analysis of A/U-rich RNA Binding Protein Using Immobilized RNA by EDC Crosslinking".PROGRESS IN BIOCHEMISTRY AND BIOPHYSICS 40.4(2013):386-391. |
入库方式： OAI收割
浏览0
下载0
收藏0
其他版本
除非特别说明，本系统中所有内容都受版权保护，并保留所有权利。