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Transformation of the L-Asparaginase II Gene to Potato Hairy Roots for Production of Recombinant Protein

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Abstract

Nowadays, transgenic plants are considered as an expression system for therapeutic recombinant proteins. One of the approaches to recombinant protein production is the introduction of the relevant genes to plant cells and produce hairy roots from these cells using the plant tissue culture methods. L-asparaginase II is one of the important therapeutic enzymes used in the treatment of cancer, especially in the treatment of acute lymphoblastic leukemia (ALL) in children. In this study, L-asparaginase II (ansB) gene was isolated from Escherichia coli YG001 and overexpressed in Solanum tuberosum. The sequence of L-asparaginase II gene was optimized according to the codon usage table of S. tuberosum and cloned into the plant expression vector pBI121. Protein, DNA, and RNA of transgenic hairy roots were investigated. It was demonstrated that the ansB gene was correctly integrated into the genomic DNA of transgenic plants. The whole protein of transgenic roots was used for the assay of L-asparaginase II activity, the higher activity of L-asparaginase II was observed in transgenic hairy roots when compared with non-transgenic ones, and the recombinant protein was detected using western blot.

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Authors and Affiliations

  1. Institute of Biotechnology, Shiraz University, Shiraz, Iran

    Azadeh Mohammadi, Ali Niazi & Farzaneh Aram

  2. Seed and Plant Certification & Registration Research Institute (SPCRI), Karaj, Iran

    Farshid Hassani

  3. Pharmaceutical Research Center, and Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran

    Younes Ghasemi

Authors
  1. Azadeh Mohammadi
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  2. Ali Niazi
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  3. Farzaneh Aram
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  4. Farshid Hassani
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  5. Younes Ghasemi
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Correspondence to Ali Niazi.

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The authors declare that they have no competing interests.

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Mohammadi, A., Niazi, A., Aram, F. et al. Transformation of the L-Asparaginase II Gene to Potato Hairy Roots for Production of Recombinant Protein. J. Crop Sci. Biotechnol. 23, 81–88 (2020). https://doi.org/10.1007/s12892-018-0030-0

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  • DOI: https://doi.org/10.1007/s12892-018-0030-0

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