Establishment of a high-throughput screening system for universal anti-hiv targets
文献类型：期刊论文
| 作者 | Yin Qi1,2; Zhuang DaoMing3; Jiang YaQin1,2; Zhao ChuanKe1,2; Zeng Xin1,2; Li ShiYou1 |
| 刊名 | Chinese science bulletin
 |
| 出版日期 | 2010-04-01 |
| 卷号 | 55期号:10页码:937-942 |
| 关键词 | Hiv-1 High-throughput screening Inhibitor Virus-cell-based assay |
| ISSN号 | 1001-6538 |
| DOI | 10.1007/s11434-009-0739-5 |
| 通讯作者 | Li shiyou(lishiyou@big.ac.cn) |
| 英文摘要 | The process of identifying novel human immunodeficiency virus 1 (hiv-1) inhibitors presents a challenge for industrial and scientific research. virus-cell-based screening approaches offer some advantages in the quest for novel inhibitors because they include multiple targets in a single screen and in some cases reveal targets not captured in biochemical assays. in this study, a high-throughput screening (hts) system for hiv-1 inhibitors was developed, which allows the simultaneous screening of all the hiv-1 targets required for replication in the cell culture. hela/cluster of differentiation 4 (cd4)/long terminal repeat (ltr) indicator cells, which stably expressed high levels of hiv receptor cd4 and contained the firefly luciferase reporter gene under the control of the hiv-1 ltr promoter, were generated. the expression of cd4 and ltr function in this cell line was validated by western blot and luciferase assay. mt2 cells, a human t-cell leukemia cell line that support high levels of hiv-1 replication, were infected with hiv-1, and then the infected mt2 cells were co-cultured with hela/cd4/ltr cells. in the optimized assay conditions, hiv-1 replication occurs rapidly in the mt2 cells, resulting in the infection of the hela/cd4/ltr cells and a significant induction of luciferase signals through ltr, which is activated by the expression of hiv-1 tat gene. the luciferase signals of hela/cd4/ltr cells co-cultured with hiv-1 infected mt2 cells were significantly stronger than the signals of noninfected hela/cd4/ltr cells (p < 0.001). the inhibitory effects of hiv-1 inhibitors (3'-azido-3'-deoxythymidine [azt], efavirenz [efv], and nevirapine [nvp]) were evaluated with this assay, and the inhibitory concentration 50% (ic(50)) values of the above three inhibitors were 58, 1.4, and 85 nmol/l, respectively, indicating that the assay provides the necessary sensitivity for identifying antiviral molecules. the z' factor had a value of 0.563, indicating this is a very robust assay. these results suggested that hiv-1 infection assay represents a novel approach to hiv-1 antiviral screening that allows for the effective execution of hts campaigns. |
| WOS关键词 | IMMUNODEFICIENCY-VIRUS TYPE-1 ; DRUG-RESISTANCE MUTATIONS ; RECEIVING HAART ; INHIBITORS ; REPLICATION ; PLASMA ; CELLS ; ASSAY ; ENHANCEMENT ; DISCOVERY |
| WOS研究方向 | Science & Technology - Other Topics |
| WOS类目 | Multidisciplinary Sciences |
| 语种 | 英语 |
| WOS记录号 | WOS:000276483000008 |
| 出版者 | SCIENCE PRESS |
| URI标识 | http://www.irgrid.ac.cn/handle/1471x/2407592 |
| 专题 | 中国科学院大学 |
| 通讯作者 | Li ShiYou |
| 作者单位 | 1.Chinese Acad Sci, Beijing Inst Genom, Beijing 100029, Peoples R China 2.Chinese Acad Sci, Grad Univ, Beijing 100049, Peoples R China 3.Acad Mil Med Sci, Inst Microbiol & Epidemiol, State Key Lab Pathogen & Biosecur, Beijing 100071, Peoples R China |
| 推荐引用方式 GB/T 7714 | Yin Qi,Zhuang DaoMing,Jiang YaQin,et al. Establishment of a high-throughput screening system for universal anti-hiv targets[J]. Chinese science bulletin,2010,55(10):937-942. |
| APA | Yin Qi,Zhuang DaoMing,Jiang YaQin,Zhao ChuanKe,Zeng Xin,&Li ShiYou.(2010).Establishment of a high-throughput screening system for universal anti-hiv targets.Chinese science bulletin,55(10),937-942. |
| MLA | Yin Qi,et al."Establishment of a high-throughput screening system for universal anti-hiv targets".Chinese science bulletin 55.10(2010):937-942. |
入库方式： iSwitch采集
来源：中国科学院大学
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。