Real-time monitoring of disintegration activity of catalytic core domain of HIV-1 integrase using molecular beacon

被引：5
|
作者：
Zhang, Da-wei [1 ,2 ]
Zhao, Ming-ming [1 ,2 ]
He, Hong-qiu [3 ]
Guo, Shun-xing [1 ,2 ]
机构：
[1] Chinese Acad Med Sci, Inst Med Plant Dev, Beijing 100193, Peoples R China
[2] Peking Union Med Coll, Beijing 100193, Peoples R China
[3] Chongqing Acad Sci & Technol, Chongqing Ctr Biomed & Med Equipment, Chongqing 401123, Peoples R China
来源：
ANALYTICAL BIOCHEMISTRY | 2013年 / 440卷 / 02期
基金：
中国国家自然科学基金;
关键词：
Molecular beacon; HIV-1; Integrase; IN-CCD; Disintegration; HUMAN-IMMUNODEFICIENCY-VIRUS; TYPE-1; INTEGRASE; IN-VITRO; DNA; PROTEIN; ASSAY; CLEAVAGE; BINDING;
D O I：
10.1016/j.ab.2013.05.032
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
HIV-1 integrase, an essential enzyme for retroviral replication, is a validated target for anti-HIV therapy development. The catalytic core domain of integrase (IN-CCD) is capable of catalyzing disintegration reaction. In this work, a hairpin-shaped disintegration substrate was designed and validated by enzyme-linked immunosorbent assay; a molecular beacon-based assay was developed for disintegration reaction of IN-CCD. Results showed that the disintegration substrate could be recognized and catalyzed by IN-CCD, and the disintegration reaction can be monitored according to the increase of fluorescent signal. The assay can be applied to real-time detection of disintegration with advantages of simplicity, high sensitivity, and excellent specificity. (C) 2013 Elsevier Inc. All rights reserved.
引用
收藏
页码：120 / 122
页数：3
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