[Establishment of a microbial assay for screening anti-human immunodeficiency virus type-1 protease inhibitors].
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.
Leng
Q
Q
Department of Virology, Institute of Medicinal Biotechnology, CAMS and PUMC, Beijing 100050.
Tao
P
P
Wang
L
L
Gao
Q
Q
D004926:Escherichia coli / Q000235:genetics*; D005818:Genetic Engineering; D016333:HIV Protease / Q000096:biosynthesis / Q000235:genetics*; D017320:HIV Protease Inhibitors / Q000494:pharmacology; D008826:Microbial Sensitivity Tests; D010957:Plasmids; D012097:Repressor Proteins / Q000096:biosynthesis / Q000235:genetics*; D013753:Tetracycline Resistance / Q000235:genetics*
D017320:HIV Protease Inhibitors; D012097:Repressor Proteins; C034811:tetracycline resistance-encoding transposon repressor protein; D016333:HIV Protease