A SimpleEscherichia coliSystem for Monitoring HIV Protease Activity: Analysis of Two Temperature-Sensitive Protease Mutants
| Autor: | Mary K. Olsen, Sue K. Rockenbach, Che-Shen C. Tomich |
|---|---|
| Rok vydání: | 1990 |
| Předmět: |
Operon
medicine.medical_treatment Genetic Vectors Immunology Mutant Gene Products gag Gene Products pol Biology medicine.disease_cause HIV Protease Virology Endopeptidases Escherichia coli medicine chemistry.chemical_classification Protease Mutagenesis Temperature Molecular biology NS2-3 protease Infectious Diseases Enzyme chemistry Biochemistry Mutation MASP1 |
| Zdroj: | AIDS Research and Human Retroviruses. 6:543-552 |
| ISSN: | 1931-8405 0889-2229 |
| DOI: | 10.1089/aid.1990.6.543 |
| Popis: | A simple Escherichia coli system has been developed for the detection of human immunodeficiency virus (HIV) protease activity. In this system, the protease sequence is placed downstream of the HIV gag polypeptide in an operon arrangement. Upon expression of the operon, gag serves as the substrate for the protease; the level of protease activity can be determined by measurement of the cleavage product of gag in cell extracts by Western immunoblotting. This system is useful in both detection of protease mutations generated by mutagenesis and in testing substrate specificity of the protease by mutagenesis of the gag sequence. Using this system, we have observed that modification of the N-terminus of HIV protease renders the enzyme temperature sensitive; the temperature sensitivity is made more pronounced by the conserved change of valine to isoleucine at residue eleven. |
| Databáze: | OpenAIRE |
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