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High-level expression of a polypeptides encoded by a large segment of the envelope gene of HIV-1 in E. coli by directed evolution

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Abstract

A random mutagenesis library of gp120-801 (a large segment of the envelope protein gene of HIV-1) was constructed using error-prone PCR and DNA shuffling methods, and one mutant of gp120-801 was selected from this library using a green fluorescent protein (GFP) fusion vector. After being cloned into pEX31b and expressed in E. coli, the expressed fusion protein reached to 15% of total bacterial proteins for the mutant but was just 1–2% for the wild type.

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Authors and Affiliations

  1. Molecular Genetics Key Laboratory, Institute of Biotechnology, Beijing, P.O. Box 130(8), P.R. China

    Yongshu Zhang, Zhihu Zhao, Cheng Cao, Chuanxuan Liu & Qingjun Ma

  2. Department of Microbiology and Immunity, HeNan Medical University, Zhengzhou, 450052, P.R. China

    Fuguang Li

Authors
  1. Yongshu Zhang
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  2. Zhihu Zhao
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  3. Fuguang Li
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  4. Cheng Cao
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  5. Chuanxuan Liu
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  6. Qingjun Ma
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Zhang, Y., Zhao, Z., Li, F. et al. High-level expression of a polypeptides encoded by a large segment of the envelope gene of HIV-1 in E. coli by directed evolution. Biotechnology Letters 22, 947–950 (2000). https://doi.org/10.1023/A:1005680622645

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  • DOI: https://doi.org/10.1023/A:1005680622645

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