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Improving the accumulation of recombinant human serum albumin (HSA) in transgenic tobacco plants by fusion with the N-terminal proline-rich domain of γ-zein (Zera)

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Abstract

Plants have long played a major role in human health as a source of herbal remedies. Recently, transgenic plants have become convenient bioreactors for the large-scale production of recombinant biopharmaceuticals at affordable prices. Manufacture of human serum albumin (HSA) in transgenic plant tissue is viable as a safe and economical alternative to the plasma fractionation method. Here, we investigated improvement of recombinant HSA accumulation in tobacco by inducing protein body formation via addition of the Zera (N-terminal proline-rich region of maize γ-zein protein) domain. Genetically modified (GM) tobacco plants were regenerated from leaf explants infected with Agrobacterium tumefaciens strains LBA4404 and GV3101 bearing pBI121-HSA or pBI121-Zera-HSA expression vectors. The T1 tobacco plantlets were examined using southern and western blotting to confirm the integration and expression of the transgenes, respectively. The confirmed T1 tobacco plantlets were further analyzed by qualitative real-time reverse transcriptase polymerase chain reaction (qRT-PCR) and enzyme-linked immunosorbent assay (ELISA). The qRT-PCR results demonstrated that rHSA gene was successfully transcribed in tobacco transformants and there was no statistically significant difference between HSA and Zera-HSA transcript levels. Despite the equivalent transcript levels, ELISA results demonstrated a nearly 4-fold increase in accumulation of HSA protein in transformed tobacco when using the Zera domain.

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Funding

Cellular and Molecular Biology Research Center at Shahid Beheshti University of Medical Sciences (Grant No. 27457) supported this research.

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Authors and Affiliations

  1. Department of Biotechnology, Faculty of Agriculture and Natural Resources, Imam Khomeini International University, Qazvin, Iran

    Behnam Sedaghati & Raheem Haddad

  2. Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran

    Behnam Sedaghati, Mojgan Bandehpour & Bahram Kazemi

  3. Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran

    Mojgan Bandehpour & Bahram Kazemi

Authors
  1. Behnam Sedaghati
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  2. Raheem Haddad
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  3. Mojgan Bandehpour
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  4. Bahram Kazemi
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Corresponding author

Correspondence to Mojgan Bandehpour.

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Editor: Pamela Weathers

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Sedaghati, B., Haddad, R., Bandehpour, M. et al. Improving the accumulation of recombinant human serum albumin (HSA) in transgenic tobacco plants by fusion with the N-terminal proline-rich domain of γ-zein (Zera). In Vitro Cell.Dev.Biol.-Plant 58, 921–930 (2022). https://doi.org/10.1007/s11627-021-10216-x

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  • DOI: https://doi.org/10.1007/s11627-021-10216-x

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