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A Novel Cytokine Derived Fusion Tag for Over- Expression of Heterologous Proteins in E. coli

Authors: S. Banerjee, A. Apte Deshpande, N. Mandi, S. Padmanabhan

Abstract:

We report a novel fusion tag for expressing recombinant proteins in E. coli. The fusion tag is the C-terminus part of the human GMCSF gene comprising 45 amino acids, which aid in over expression of otherwise non expressible genes. Expression of hIFN a2b with this fusion tag also escapes the requirement of rare codons for expression. This is also a first report of a small fusion tag of human origin having affinity to heparin sepharose column facilitating the purification of fusion protein.

Keywords: fusion tag, bacterial expression, rare codons, human GMCSF

Digital Object Identifier (DOI): doi.org/10.5281/zenodo.1078231

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References:


[1] R G Harrison, "Expression of soluble heterologous proteins via fusion with NusA protein", Innovations, vol 11, pp 4-7, 2000.
[2] D M Valeria, S Gunter, B Stephanie, D M Ario, "The solubility and stability of recombinant proteins are increased by their fusion to NusA.", Biochem. Biophys. Res. Commun., vol 322, pp 766-771, 2004.
[3] L D Cabrita, W Dai, S P Bottomley, "A family of E. coli expression vectors for laboratory scale and high throughput soluble protein production", BMC biotechnology, vol 6, pp 12-19, 2006.
[4] MC Smith, TC Furman, TD Ingolia, and C Pidgeon , "Chelatingpeptideimmobilized metal ion affinity chromatography. A new concept in affinity chromatography for recombinant proteins", J. Biol. Chem., vol 263, pp 7211-7215, 1988.
[5] E R LaVallie, E A DiBlasio, S Kovacic, K L Grant, P F Schendel and J M McCoy, "A Thioredoxin Gene Fusion Expression System That Circumvents Inclusion Body Formation in the E. coli Cytoplasm", Biotechnol., vol 11, pp 187-193, 1993.
[6] D B Smith, K S Johnson, "Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase", Gene, vol 67, pp 31-40, 1988.
[7] D C Guan, P Li, P D Riggs, H Inouye, "Vectors that facilitate the expression and purification of foreign peptides in Escherichia coli by fusion to maltose-binding protein", Gene, vol 67, pp 21-30, 1988.
[8] G D Davis, C Elisee, D M Newham and R G Harrison, "New fusion protein systems designed to give soluble expression in Escherichia coli", Biotechnol. Bioeng. Vol 65, pp 382-388, 1999.
[9] Y Kanakura, S A Cannistra, C B Brown, M Nakamura, G F Seelig, W W Prosise et. al., "Identification of functionally distinct domains of human granulocyte-macrophage colony-stimulating factor using monoclonal antibodies", Blood, vol 77, pp 1033-1043, 1991.
[10] A Sebollela, T C Cagliaris, G S C S Limaverde, A chapeaurouge, M H F Sorgine, T C Sampaio et. al., " Heparin binding sites in granulocyte macrophage colony stimulating factor", J. Biol. Chem., vol 280, pp 31049-31956, 2005.
[11] H W Lee, J H Joo, S Kang, I S Song, J B Kwon, M H Han and D S Na, "Expression of human interleukin-2 from native and synthetic genes in E. coli: No correlation between major codon bias and high level expression", Biotechnol. Lett., vol 14, pp 653-658, 1992.
[12] D S Waugh, "Making the most of affinity tags", Trends in biotech, vol 23, pp 316-320, 2005.
[13] J J Olivares-Trejo, J G Bueno-Martínez, G Guarneros,and J Hernández- Sánchez, "The pair of arginine codons AGA AGG close to the initiation codon of the lambda int gene inhibits cell growth and protein synthesis by accumulating peptidyl-tRNAArg4", Molecular Micribiology, vol 49, pp 1043-1049, 2003.
[14] O L Garcia, B Gonzalez, A Menendez, A E Sosa, J R Fernandez, H Santana and N Meneses, "The argU gene product enhances expression of the recombinant a2- Interferon in Escherichia coli", Annals New York Academy of Sciences, vol 782, pp 79-86, 1996.
[15] X Wu, H Jornvall, K D Berndt and U Oppermann, "Codon optimization reveals critical factors for high level expression of two rare codon genes in Escherichia coli: RNA stability and secondary structure", Biochem. Biophys. Res. Commun., vol 313, pp 89-96, 2004.
[16] S Hatano, H Asano, H Nagai, T Murate, N Mori, K Kawashima et. Al., " In vitro effect of recombinant human IL-11 in human malignant lymphoma cells", J. Clin. Exp. Hematopathol., vol 42, pp 55-60, 20.