title: [Establishment of a microbial assay for screening anti-human immunodeficiency virus type-1 protease inhibitors].
abstract: 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.
authors.affiliation: Department of Virology, Institute of Medicinal Biotechnology, CAMS and PUMC, Beijing 100050.
authors.forename: Q
authors.initials: Q
authors.lastname: Leng
authors.forename: P
authors.initials: P
authors.lastname: Tao
authors.forename: L
authors.initials: L
authors.lastname: Wang
authors.forename: Q
authors.initials: Q
authors.lastname: Gao
mesh_terms: 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*
chemical_list: D017320:HIV Protease Inhibitors; D012097:Repressor Proteins; C034811:tetracycline resistance-encoding transposon repressor protein; D016333:HIV Protease