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Journal of Virology, March 2002, p. 3031-3037, Vol. 76, No. 6
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.6.3031-3037.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Construction of a Human Immunodeficiency Virus Type 1 (HIV-1) Library Containing Random Combinations of Amino Acid Substitutions in the HIV-1 Protease due to Resistance by Protease Inhibitors

Keisuke Yusa,1* Wei Song,1 Matthias Bartelmann,1,2 and Shinji Harada1,3

Department of Medical Virology, Kumamoto University School of Medicine,1 Center for AIDS Research, Kumamoto University, Kumamoto 860-0811,3 Japanese Foundation for AIDS Prevention, Minato-ku, Tokyo 105-0001, Japan2

Received 4 September 2001/ Accepted 14 December 2001

Human immunodeficiency virus type 1 (HIV-1) heterogeneity contributes to the emergence of drug-resistant virus, escape from host defense systems, and/or conversion of the cellular tropism. To establish an in vitro system to address a heterogeneous virus population, we constructed a library of HIV-1 molecular clones containing a set of random combinations of zero to 11 amino acid substitutions associated with resistance to protease inhibitors by the HIV-1 protease. The complexity (2.1 x 105) of the HIV-1 library pNG-PRL was large enough to cover all of the possible combinations of zero to 11 amino acid substitutions (a total of 4,096 substitutions possible). The T-cell line MT-2 was infected with the HIV-1 library, and resistant viruses were selected after treatment by the protease inhibitor ritonavir (0.03 to 0.30 µM). The viruses that contained three to eight amino acid substitutions could be selected within 2 weeks. These results demonstrate that this HIV-1 library could serve as an alternative in vitro system to analyze the emergence of drug resistance and to evaluate the antiviral activity of novel compounds against multidrug-resistant viruses.

* Corresponding author. Mailing address: Department of Medical Virology, Kumamoto University School of Medicine, 2-2-1 Honjo, Kumamoto 860-0811, Japan. Phone: 81-96-373-5129. Fax: 81-96-373-5132. E-mail: yusak{at}kaiju.medic.kumamoto-u.ac.jp.

Journal of Virology, March 2002, p. 3031-3037, Vol. 76, No. 6
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.6.3031-3037.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Yusa, K., Maeda, Y., Fujioka, A., Monde, K., Harada, S. (2005). Isolation of TAK-779-resistant HIV-1 from an R5 HIV-1 GP120 V3 Loop Library. J. Biol. Chem. 280: 30083-30090 [Abstract] [Full Text]  


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