Construction of a cell-based high-flux assay for the rev protein of HIV-1.
The Rev of HIV-1 is essential for the replication of the viruses and is therefore a very attractive target for the development of antiviral drugs. To establish a cell-based high-flux assay system for random screening of Rev inhibitors, cells carrying both the Rev-expressing gene and a Rev-inducible SeAP gene were generated by permanent transfection. SeAP produced by these cells was 5-10-fold higher than that synthesized by cells not carrying the Rev gene. Northern blot analysis demonstrated that the increase in SeAP was due mainly to an increase in SeAP transcripts, indicating the effect of Rev proteins synthesized by the transfected cells. The assay system reported in this study should be useful for screening novel Rev inhibitors.
Tang
R Y
RY
College of Life Science, National Yang-Ming University, Taipei, Taiwan, R.O.C.
Su
Y
Y
D000469:Alkaline Phosphatase / Q000737:chemistry / Q000235:genetics; D019380:Anti-HIV Agents / Q000302:isolation & purification*; D015139:Blotting, Southern; D002460:Cell Line; D004274:DNA, Recombinant / Q000096:biosynthesis / Q000187:drug effects; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D005786:Gene Expression Regulation / Q000187:drug effects; D015694:Gene Products, rev / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics*; D017930:Genes, Reporter; D015497:HIV-1 / Q000737:chemistry* / Q000187:drug effects / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000737:chemistry; D014158:Transcription, Genetic; D014162:Transfection; D054321:rev Gene Products, Human Immunodeficiency Virus
D019380:Anti-HIV Agents; D004274:DNA, Recombinant; D015694:Gene Products, rev; D012333:RNA, Messenger; D054321:rev Gene Products, Human Immunodeficiency Virus; D000469:Alkaline Phosphatase