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Production of bovine interferon-gamma in transgenic tobacco plants

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Abstract

The inbreed line of Nicotiana tabacum L. plants harboring a heterologous gene of bovine interferon-gamma sIFNG under the control of constitutive 35S CaMV promoter was created by Agrobacterium-mediated genetic transformation. The transformation of leaf discs yielded six independent transgenic plants (T0 generation). All six transformants demonstrated presence of sIFNG insertion and had a normal phenotype. Self-pollination of T0 plants provided six transgenic families. All families but one inherited sIFNG insertion in T1 progeny and were used for subsequent selection. Offsprings of selfing were tested for the presence and expression of sIFNG gene and the synthesis of bovine interferon-gamma protein. We identified homozygous sIFNG plant of Inter311.2 family in T1 generation, which was laid the founder of the inbreed line of transgenic sIFNG tobacco. The line demonstrated stable inheritance and expression of the transgene insertion and presence of a heterologous interferon-gamma protein in the plant tissue up to the fourth generation of transgenic plants tested. Antiviral and immunomodulatory activities of plant-produced interferon-gamma upon bovine cell cultures and laboratory animals (mice) were observed. Created tobacco plants may be used as the bioreactors for production of bovine interferon-gamma for veterinary.

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Abbreviations

IFN:

Interferon

IgG:

Class G immunoglobulin

MS0:

Murashige and Skoog basal medium

PCR:

Polymerase chain reaction

RT-PCR:

Reverse transcription-polymerase chain reaction

sIFNG :

Synthetic bovine interferon-gamma gene

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Acknowledgments

Authors would like to thank Dr. L.T. Khodjaova for the help with plasmid construction and plant transformation, and Dr. A.E. Zobnina (nee Gradoboeva) for assistance with testing of recombinant IFN-γ in cell culture. The research was supported by research resource center “Molecular and cell technologies” of St. Petersburg State University. Financial support from Ministry of Education and Science of Russian Federation (state contract 02.740.11.0768) and St. Petersburg State University (projects 1.39.1056.2012 and 1.38.229.2014) are greatly appreciated.

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Correspondence to V. V. Yemelyanov.

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Burlakovskiy, M.S., Saveleva, N.V., Yemelyanov, V.V. et al. Production of bovine interferon-gamma in transgenic tobacco plants. Plant Cell Tiss Organ Cult 122, 685–697 (2015). https://doi.org/10.1007/s11240-015-0802-7

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Keywords

  • Tobacco
  • Interferon-gamma
  • Immunomodulator
  • Veterinary
  • Transgenic plants
  • Heterologous expression