Related Experiment Video
Updated: Sep 27, 2026
Rapid Screening of HIV Reverse Transcriptase and Integrase Inhibitors
Published on: April 9, 2014
[Establishment of a microbial assay for screening anti-human immunodeficiency virus type-1 protease inhibitors]
1Department of Virology, Institute of Medicinal Biotechnology, CAMS and PUMC, Beijing 100050.
Objective:
This study was to establish a microbial assay of human immunodeficiency virus type-1 protease (HIV-1 PR) activity for screening anti-HIV PR inhibitors.
Methods:
A 24 bp synthetic oligonucleotide fragment that encodes the HIV-1 PR recognition sequence was inserted into the tetr gene of pBR322 (mtetr). Escherichia coli containing HIV-1 PR expression vector-pPOLO was transformed with pACYC184M containing modified mtetr gene. The transformant could express both HIV-1 PR and the modified Tet protein.
Results:
The growth of engineered E. coli was prevented in the presence of tetracycline because the resistance Tet protein was degraded by HIV-1 PR. However inhibition of the HIV-1 PR restored tetracycline resistance. 31 chemical synthetic compounds were tested by the microbial assay.
Conclusions:
A microbial assay method of HIV-1 PR activity was established through a engineered E. coli. 5 mumol/L saqunavir-a special HIV-1 PR inhibitor showed inhibitory effect on the engineered E. coli. That means this model could be used as a initial screening model for anti-HIV PR agents.
