以p-TEFb为靶点抗HIV药物高通量筛选模型建立及在中药初筛中的应用  

Establishment of high throughput screening model for anti-HIV drugs with p-TEFb as target and its application in screening Chinese meteria medica

作　　者：李芬[1,2,3] 李玉会 吴秀丽[1,2] 胡岩岩[1,2] 马瑜[1,2] 贾盼盼[1] 岳晓杰[1] 李明军 

机构地区：[1]河南师范大学生命科学学院,河南新乡453007 [2]资源微生物与功能分子河南省高校重点实验室培育基地,河南新乡453007 [3]河南省高校道地中药材保育及利用工程技术研究中心,河南新乡453007 [4]河南省项城市中医院,河南项城466200

出　　处：《中草药》2014年第5期679-685,共7页Chinese Traditional and Herbal Drugs

基　　金：河南省重点科技攻关项目(092102310026);河南省高校科技创新团队支持计划资助项目(13IRTSTHN009)

摘　　要：目的建立以正向转录延伸因子(p-TEFb)为靶点的抗HIV药物快速高通量筛选模型,并运用此模型筛选抗艾滋病中药。方法构建BD-Tat和AD-CyclinT1融合的酵母双杂交质粒,分别转入AH109和Y187,经接合实验获得二倍体菌株并对其进行毒性检测、自激活实验和报告基因表达检测,建立基于酵母双杂交的筛选模型。结果运用此系统筛选具有提高机体免疫功效的20种中药,获阳性中药2种,其抗HIV活性已被体外抑制HIV实验证实。结论该筛选模型可成功用于抗艾滋病化合物和中药的高效筛选,得到的2种阳性中药狗脊和黄连,值得进一步进行干扰HIV Tat和CyclinT1互作的有效成分分离研究。Objective To establish a rapid high-throughput screening model with positvive transcription elongation factor （p-TEFb） as target for screening the inhibitors of HIV. Methods Double cross plasmid of BD-Tat and AD-CyclinT 1 was established and transformed into yeast AH 109 and Y 187, respectively. AH 109/pGBKT7-Tat was mated with Y 187/pGADT7-CyclinT 1 and the diploids were subjected to autoactivation, toxicity test, and reporter gene assay. The screening model based on double hybrid system was established. Results Two out of 20 kinds of Chinese materia medica possessing immunity-enhancing effects were identified as primary hits, the anti-HIV activity of these two positive drugs was already demonstrated by other researchers through detecting the suppression effects on HIV duplication in vitro. Conclusion The system established in this paper can be used for rapid high-throughput screening anti-HIV chemicals or herbs. Further research for the obtained two positive Chinese materia medica, Cibotii Rhizoma and Coptidis Rhizom should be carded out to isolate the effective comoonent for disrupting the interaction between HIV Tat and CyclinT1.

关 键 词：正向转录延伸因子 靶点 HIV 高通量筛选模型 狗脊 黄连 

分 类 号：R285.5[医药卫生—中药学]