Expression and purification of HIV-1 protease and the establishment of a method for protease inhibitor screening

Virologica Sinica

Volumn 21, Issue 2, 2006, Pages 126-130

  Wang, Yun Hua ^a,c   Wang, Rui Rui ^a,c   Yang, Liu Meng ^a   Li, Jian Feng ^b   Xu, Wei Ming ^b   Zheng, Yong Tang ^a  

^a KUNMING INSTITUTE OF ZOOLOGY   (China)

^b INSTITUTE OF MEDICAL BIOLOGY   (China)

^c UNIVERSITY OF CHINESE ACADEMY OF SCIENCES   (China)

Author keywords

Expression; HIV 1; Inhibitors; Protease; Purification

Indexed keywords

BACTERIAL PROTEIN; BUFFER; DILUENT; INDINAVIR; PROTEINASE; PROTEINASE INHIBITOR; RECOMBINANT ENZYME; TRITON X 100; UREA; VIRUS RNA;

ANALYTIC METHOD; ARTICLE; CELL CULTURE; CELL INCLUSION; CONTROLLED STUDY; CULTURE MEDIUM; DILUTION; DISSOLUTION; DRUG SCREENING; ENZYME ACTIVITY; ESCHERICHIA COLI; EXPRESSION VECTOR; FLUORESCENCE ANALYSIS; FLUORESCENCE RESONANCE ENERGY TRANSFER; GENE AMPLIFICATION; GENE INSERTION; GENE SEQUENCE; GENETIC CODE; GENETIC TRANSFORMATION; HUMAN IMMUNODEFICIENCY VIRUS 1; MOLECULAR CLONING; NONHUMAN; PROTEIN EXPRESSION; PROTEIN PURIFICATION; REVERSE TRANSCRIPTION POLYMERASE CHAIN REACTION; START CODON; STOP CODON; ULTRAFILTRATION;

BACTERIA (MICROORGANISMS); ESCHERICHIA COLI; HUMAN IMMUNODEFICIENCY VIRUS 1;

EID: 33645538059     PISSN: 10035125     EISSN: None     Source Type: Journal    
DOI: None     Document Type: Article

References (10)

1. Daar E S Richman D D. Confronting the emergence of drug-resistant HIV type 1: Impact of antiretroviral therapy on individual and population resistance [J]. AIDS Res Hum Retroviruses, 2005, 21(5): 343-57.
2. Clavel F, Hance A J. HIV drug resistance [J]. N Engl J Med, 2004, 350 (10): 1023-1035.
3. Zheng Y T. In vitro screening and assays for anti-HIV agents [J]. Chin J New Drugs, 2002, 11 (8): 596-600.
4. Tomasselli A G, Heinrikson R L. Targeting the HIV protease in AIDS therapy: A current clinical perspective [J]. Biochem Biophys Acta, 2000, 1477:189-214.
5. Matayoshi E D, Wang G T, Krafft G A, et al. Novel fluorogenic sustrate for assaying retroviral protease by resonance energy transfer [J]. Science, 1990, 247 (4945): 954-958.
6. Hickman D, Vasavanonda S, Nequist G, et al. Estimation of Serum-Free 50-Percent Inhibitory Concentrations for Human Immunodeficiency Virus Protease Inhibitors Lopinavir and Ritonavir [J]. Antimicrob. Agents Chemother, 2004, 48 (8): 2911-2917.
7. Yan X J, Su C Z, Ji C H et al. High level expression of HIV-1 Pr in Escherichia Coli [J]. Chin J Microbiol Immunol, 1994, 14 (2): 89-92.
8. Leuthardt A, Roesel J L. Cloning, expression and purification of a recombinant poly-histidine-linked HIV-1 protease [J]. FEBS Lett, 1993, 326 (1-3): 275-280.
9. Vickrey J F, Logsdon B C, Proteasa G, et al. HIV-1 protease variants from 100-fold drug resistant clinical isolates: Expression, purification, and crystallization [J]. Protein Expr Purif, 2003, 28: 165-172.
10. Wan M, Loh B N. Expression and purification of active form of HIV-1 protease from E.coli [J]. Biochem Mol Biol Int, 1995, 35(4): 899-912