Novel bacteriological assay for detection of potential antiviral agents.
A prototype assay for the initial screening of potential antiviral agents that uses bacterial growth on selective media is described. The human immunodeficiency virus (HIV) protease recognition sequence was inserted into the tetracycline resistance (Tet) protein encoded by plasmid pBR322 of Escherichia coli. Expression of both the HIV protease and the modified Tet protein prevented growth in the presence of tetracycline. However, inhibition of the HIV protease restored tetracycline resistance. Thus, potential HIV protease inhibitors can be identified by their ability to confer tetracycline resistance to this bacterial strain. The assay is simple, rapid, and inexpensive, and this concept can be applied to the search for inhibitors of other viral proteases.
Block
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Department of Microbiology and Immunology, Thomas Jefferson University, Philadelphia, Pennsylvania 19107.
Grafstrom
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D000998:Antiviral Agents / Q000494:pharmacology*; D001483:Base Sequence; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D006678:HIV / Q000187:drug effects / Q000201:enzymology; D016333:HIV Protease / Q000378:metabolism; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D010436:Pepstatins / Q000494:pharmacology; D010957:Plasmids; D012097:Repressor Proteins / Q000138:chemical synthesis / Q000737:chemistry; D015183:Restriction Mapping; D013752:Tetracycline / Q000494:pharmacology; D013753:Tetracycline Resistance / Q000235:genetics
D000998:Antiviral Agents; D010436:Pepstatins; D012097:Repressor Proteins; C034811:tetracycline resistance-encoding transposon repressor protein; C014195:Streptomyces pepsin inhibitor; D016333:HIV Protease; D013752:Tetracycline; C031375:pepstatin