Development of Wolffia arrhiza as a Producer for Recombinant Human Granulocyte Colony-Stimulating Factor

Frontiers in Chemistry
Pavel KhvatkovSergey Dolgov

Abstract

To date, the expression of recombinant proteins in transgenic plants is becoming a powerful alternative to classical expression methods. Special efforts are directed to the development of contained cultivation systems based on cell culture or rhyzosecretion, which reliably prevents the heterologous DNA releasing into the environment. A promising object for the development of such systems is the tiny aquatic plant of Wolffia arrhiza, which can be used as a dipped culture in bioreactors. Herein we have expressed the human granulocyte colony-stimulating factor (hG-CSF) in nuclear-transformed Wolffia. The nucleotide sequence of hG-CSF was optimized for expression in Wolffia and cloned into the vector pCamGCSF downstream of double CaMV 35S promoter. Wolffia plants were successfully transformed and 34 independent transgenic lines with hG-CSF gene were obtained, PCR and Southern blot analysis confirmed the transgenic origin of these lines. Western blot analysis revealed accumulation of the target protein in 33 transgenic lines. Quantitative ELISA of protein extracts from these lines showed hG-CSF accumulation up to 35.5 mg/kg of Wolffia fresh weight (0.194% of total soluble protein). This relatively high yield holds promise for the de...Continue Reading

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Citations

Sep 26, 2020·Frontiers in Immunology·P P M HeenatigalaHongwei Hou
Jan 17, 2021·Biomolecules·Gui-Li YangAi-Juan Tan
May 1, 2021·3 Biotech·Pavel KhvatkovSergey Dolgov
Aug 22, 2021·BMC Plant Biology·Jingjing YangHongwei Hou

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Datasets Mentioned

BETA
DB00099
AAA33897.1

Methods Mentioned

BETA
transgenic
PCR
light microscopy
electrophoresis
ELISA
phosphotransferase

Software Mentioned

Wolffia
StatSoft
Gene2Oligo
Gene Composer
Statistica

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