Single-step purification of recombinant wild-type and mutant HIV-1 reverse transcriptase

Protein Expression and Purification

Volumn 7, Issue 1, 1996, Pages 27-32

  Fletcher, Ronald S ^a   Holleschak, Gyula ^a   Nagy, Eva ^a   Arion, Dominique ^a   Borkow, Gadi ^a   Gu, Zhengxian ^a   Wainberg, Mark A ^a   Parniak, Michael A ^a  

^a MCGILL UNIVERSITY   (Canada)

Author keywords

[No Author keywords available]

Indexed keywords

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

EID: 0030087537     PISSN: 10465928     EISSN: None     Source Type: Journal    
DOI: 10.1006/prep.1996.0004     Document Type: Article

References (19)

1. 1. Mizrahi, V., Lazarus, G. M., Miles, L. M., Meyers, C. A., and Debouck, C. (1989) Recombinant HIV-1 reverse transcriptase: Purification, primary structure and polymerase/ribonuclease H activities. Arch. Biochem. Biophys. 273, 347-358.
2. 2. Deibel, M. R., Jr., McQuade, T. J., Brunner, D. P., and Tarpley, W. G. (1990) Denaturation/refolding of purified HIV reverse transcriptase yields monomeric enzyme with high enzymatic activity. AIDS Res. Hum. Retrovir. 6, 329-340.
3. 3. Clark, P. K., Ferris, A. L., Miller, D. A., Hizi, A., Kim, K.-W., Deringer-Boyer, S. M., Mellini, M. L., Clark, A. D., Jr., Arnold, G. F., Lebherz, W. B., III, Arnold, E., Muschik, G. M., and Hughes, S. H. (1990) HIV-1 revese transcriptase purified from a recombinant strain of Escherichia coli. AIDS Res. Hum. Retrovir. 6, 753-764.
4. 4. Chattopadhyay, D., Einspahr, H. M., Brunner, D. P., Strakalaitis, N. A., Tarpley, W. G., and Deibel, M. R., Jr. (1992) Resolution of the microheterogeneity associated with recombinant HIV-1 heterodimeric reverse transcriptase. Protein Expression Purif. 3, 151-159.
5. 5. Bavand, M. R., Wagner, R., and Richmond, T. J. (1993) HIV-1 reverse transcriptase: Polymerization properties of the p51 homodimer compared to the p66/p51 heterodimer. Biochemistry 32, 10543-10552.
6. 6. Becerra, S. P., Kumar, A., and Wilson, S. H. (1993) Expression of polypeptides of human immunodeficiency virus 1 reverse transcriptase in Escherichia coli. Protein Expression Purif. 4, 187-199.
7. 7. Starnes, M., and Cheng, Y. (1989) Human immunodeficiency virus reverse transcriptase-associated RNase H activity. J. Biol. Chem. 264, 7073-7077.
8. 8. Gu, Z., Fletcher, R. S., Arts, E. J., Wainberg, M. A., and Parniak, M. A. (1994) The K65R mutant reverse transcriptase of HIV-1 cross-resistant to 2′,3′-dideoxycytidine, 2′,3′-dideoxy-3′-thiacytidine, and 2′,3′-dideoxyinosine shows reduced sensitivity to specific dideoxynucleoside triphosphate inhibitors in vitro. J. Biol. Chem. 269, 28118-28122.
9. 9. Gu, Z., Arts, E. J., Parniak, M. A., and Wainberg, M. A. (1995) Mutated K65R recombinant reverse transcriptase of human immunodeficiency virus type 1 shows diminished chain termination in the presence of 2′,3′-dideoxycytidine triphosphate and other drugs. Proc. Natl. Acad. Sci. USA 92, 2760-2764.
10. 10. Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, 2nd. ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
11. 11. Li, X., Amandoron, E., Wainberg, M. A., and Parniak, M. A. (1993) Generation and characterization of murine monoclonal antibodies reactive against N-terminal and other regions of HIV-1 reverse transcriptase. J. Med. Virol. 39, 251-259.
12. 12. Muller, B., Restle, T., Kuhnel, H., and Goody, R. S. (1991) Expression of the heterodimeric form of human immunodeficiency virus type 2 reverse transcriptase in Escherichia coli and characterization of the enzyme. J. Biol. Chem. 266, 14709-14713.
13. 13. Hizi, A., McGill, C., and Hughes, S. H. (1988) Expression of soluble, enzymatically active, human immunodeficiency virus reverse transcriptase in Escherichia coli and analysis of mutants. Proc. Natl. Acad. Sci. USA 85, 1218-1222.
14. 14. LeGrice, S. F. J., Naas, T., Wohlgensinger, B., and Schatz, O. (1991) Subunit-selective mutagenesis indicates minimal polymerase activity in heterodimer-associated p51 HIV-1 reverse transcriptase. EMBO J. 10, 3905-3911.
15. 15. Hostomsky, Z., Hostomska, Z., Fu, T.-B., and Taylor, J. (1992) Reverse transcriptase of human immunodeficiency virus type 1: Functionality of the subunits of the heterodimer in DNA synthesis. J. Virol. 66, 3179-3182.
16. 16. Thimmig, R. L., and McHenry, C. S. (1993) Human immunodeficiency virus reverse transcriptase: Expression in Escherichia coli, purification, and characterization of a functionally and structurally asymmetric dimeric polymerase. J. Biol. Chem. 268, 16528-16536.
17. 17. D'Aquila, R. T., and Summers, W. C. (1989) HIV-1 reverse transcriptase/ribonuclease H: High level expression in Escherichia coli from a plasmid constructed using the polymerase chain reaction. J. Acquired Immune Defic. Syndr. 2, 579-587.
18. 18. Tisdale, M., Ertl, P., Larder, B., Purifoy, D. J. M., Darby, G., and Powell, K. (1988) Characterization of the human immunodeficiency virus type 1 reverse transcriptase by using monoclonal antibodies: Role of the C-terminus in antibody reactivity and enzyme function. J. Virol. 62, 3662-3667.
19. 19. Lederer, H., Schatz, O., May, R., Crespi, H., Darlix, J.-L., Le-Grice, S. F. J., and Heumann, H. (1992) Domain structure of the human immunodeficiency virus reverse transcriptase. EMBO J. 11, 1131-1139.