A genetic screen for the isolation and characterization of site-specific proteases.
Site-specific proteolysis is an important regulatory mechanism in basic cellular and viral processes. Using the protease of the HIV as a model, a genetic system has been developed for the isolation and characterization of site-specific proteases. The system utilizes the well defined bacteriophage lambda regulatory circuit where the viral repressor, cI, is specifically cleaved to initiate the lysogenic-to-lytic switch. The model system is rapid, highly specific, and demonstrates the ability to isolate and characterize enzymes of limited expression or activity. In addition, the system has a significant potential for the selection of clinically relevant mutant enzymes and in the development of anti-protease therapeutics.
Sices
H J
HJ
Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Building 4-Room 133, 9000 Rockville Pike, Bethesda, MD 20892, USA.
Kristie
T M
TM
D010582:Bacteriophage lambda; D003001:Cloning, Molecular / Q000379:methods*; D004268:DNA-Binding Proteins; D010450:Endopeptidases / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D016333:HIV Protease / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D017320:HIV Protease Inhibitors / Q000494:pharmacology; D008242:Lysogeny; D012097:Repressor Proteins; D012641:Selection, Genetic; D013379:Substrate Specificity; D014764:Viral Proteins; D054334:Viral Regulatory and Accessory Proteins; D014775:Virus Activation
D004268:DNA-Binding Proteins; D017320:HIV Protease Inhibitors; D012097:Repressor Proteins; D014764:Viral Proteins; D054334:Viral Regulatory and Accessory Proteins; C016649:phage repressor proteins; D010450:Endopeptidases; D016333:HIV Protease