Efficient transient expression of human GM-CSF protein in Nicotiana benthamiana using potato virus X vector

Applied Microbiology and Biotechnology
Fengyong ZhouYun J Zhu

Abstract

The human granulocyte macrophage colony-stimulating factor (GM-CSF) is a glycoprotein with important clinical applications for the treatment of neutropenia and aplastic anemia and reducing infections associated with bone marrow transplants. We evaluated the potential for using a potato virus X (PVX) viral vector system for efficient expression of the biologically functional GM-CSF protein in Nicotiana benthamiana leaves. The GM-CSF gene was cloned into PVX viral expression vector, driven with the CaMV 35S promoter. Gene transfer was accomplished by inoculating N. benthamiana leaves with the plasmid DNA of PVX vector containing the GM-CSF gene. The expression level of the recombinant GM-CSF protein was determined with ELISA and its size was confirmed by Western blot analysis. The results showed that: (1) leaf age significantly affects GM-CSF protein concentration with younger leaves accumulating 19.8 mg g(-1) soluble protein which is 2.6 times the concentration in older leaves, (2) recombinant protein accumulation within a given leaf declined slightly over time but was not significantly different between 7 and 11 days post-inoculation (dpi), and (3) the two leaves immediately above the inoculated leaves play an important role fo...Continue Reading

References

Nov 1, 1991·International Journal of Cell Cloning·K M Sakamoto, J C Gasson
Sep 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·M A CantrellS Gillis
Jul 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·F LeeD Rennick
Oct 1, 1995·The Journal of Infectious Diseases·D C DaleS Nelson
Jun 1, 1995·The Plant Journal : for Cell and Molecular Biology·D C BaulcombeS Santa Cruz
Apr 16, 1996·Proceedings of the National Academy of Sciences of the United States of America·S GerondakisR J Grumont
Mar 14, 1998·Advances in Experimental Medicine and Biology·P Van den SteenG Opdenakker
Feb 24, 2001·Molecular Plant-microbe Interactions : MPMI·H SaitohR Terauchi
Jun 4, 2002·Nature Biotechnology·Allison C MalloryVicki B Vance
Aug 14, 2003·Journal of Virology·Sophia BiemeltMartin Müller
Nov 20, 2003·Trends in Biotechnology·Richard M TwymanRainer Fischer
Feb 14, 2004·Methods : a Companion to Methods in Enzymology·Birgit WagnerHeimo Breiteneder
Jul 19, 2005·Transgenic Research·Ming-Li WangHenrik H Albert

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Citations

Dec 30, 2009·Journal of Interferon & Cytokine Research : the Official Journal of the International Society for Interferon and Cytokine Research·Anna Góra-SochackaAgnieszka Sirko
Nov 18, 2009·Biochemistry. Biokhimii︠a︡·A S ZverevaJ G Atabekov
Jul 13, 2011·International Journal of Molecular Sciences·Agnieszka SirkoPatrycja Redkiewicz
Nov 8, 2015·Biochemistry. Biokhimii︠a︡·E V PutlyaevJ G Atabekov
Jan 1, 2016·Applied Biochemistry and Microbiology·M S BurlakovskiyL A Lutova

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