Preparation and identification of anti-transforming growth factor beta 1 U1 small nuclear RNA chimeric ribozyme in vitro
文献类型：期刊论文
| 作者 | Lin, JS; Song, YH; Kong, XJ; Li, B; Liu, NZ; Wu, XL; Jin, YX |
| 刊名 | WORLD JOURNAL OF GASTROENTEROLOGY
 |
| 出版日期 | 2003 |
| 卷号 | 9期号:3页码:572-577 |
| 通讯作者 | Jin, YX (reprint author), Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Biochem & Cell Biol, State Key Lab Mol Biol, Shanghai 200031, Peoples R China.,yxjin@sunm.shcnc.ac.cn |
| 英文摘要 | AIM: To study the preparation and cleavage activity of anti-transforming growth factor (TGF)beta1 U1 small nuclear (sn) RNA chimeric hammerhead ribozymes in vitro. METHODS: TGF(31 partial gene fragment was cloned into T-vector at the downstream of T7 promoter. (32)p-labeled TGFbeta1 partial transcripts as target RNA were transcribed in vitro and purified by denaturing polyacrylamide gel electrophoresis (PAGE). Anti-TGF01 ribozymes were designed by computer, then synthetic ribozyme fragments were cloned into the U1 ribozyme vector pZeoU1EcoSpe containing U1 snRNA promoter/enhancer and terminator. (32)p-labeled U1 snRNA chimeric ribozyme transcripts were gel-purified, incubated with target-RNAs at different conditions and autoradiographed after running denaturing PAGE. RESULTS: Active U1snRNA chimeric ribozyme (U1Rz803) had the best cleavage activity at 50 C; at 37 C, it was active, K-m=34.48 nmol/L, K-cat= 0.14 min(-1); while the point mutant ribozyme U1Rz803(m) had no cleavage activity, so these indicated the design of U1Rz803 was correct. CONCLUSION: U1Rz803 prepared in this study possessed the perfect specific catalytic cleavage activity. These results indicate U1 snRNA chimeric ribozyme U1Rz803 may suppress the expression of TGFbeta1 in vivo, therefore it may provide a new avenue for the treatment of liver fibrosis in the future. |
| 学科主题 | Gastroenterology & Hepatology |
| 类目[WOS] | Gastroenterology & Hepatology |
| 关键词[WOS] | RAT-LIVER FIBROSIS ; HAMMERHEAD RIBOZYME ; MESSENGER-RNA ; HEPATITIS-B ; IN-VITRO ; MOLECULAR REGULATION ; GENE-THERAPY ; CELLS ; FIBROGENESIS ; EXPRESSION |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000181745400040 |
| 版本 | 出版稿 |
| 源URL | [http://202.127.25.143/handle/331003/2270]  |
| 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 | Lin, JS,Song, YH,Kong, XJ,et al. Preparation and identification of anti-transforming growth factor beta 1 U1 small nuclear RNA chimeric ribozyme in vitro[J]. WORLD JOURNAL OF GASTROENTEROLOGY,2003,9(3):572-577. |
| APA | Lin, JS.,Song, YH.,Kong, XJ.,Li, B.,Liu, NZ.,...&Jin, YX.(2003).Preparation and identification of anti-transforming growth factor beta 1 U1 small nuclear RNA chimeric ribozyme in vitro.WORLD JOURNAL OF GASTROENTEROLOGY,9(3),572-577. |
| MLA | Lin, JS,et al."Preparation and identification of anti-transforming growth factor beta 1 U1 small nuclear RNA chimeric ribozyme in vitro".WORLD JOURNAL OF GASTROENTEROLOGY 9.3(2003):572-577. |
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