A Novel Cytokine Derived Fusion Tag for Over- Expression of Heterologous Proteins in E. coli
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.
[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.
[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.
@article{"International Journal of Biological, Life and Agricultural Sciences:60978", author = "S. Banerjee and A. Apte Deshpande and N. Mandi and S. Padmanabhan", title = "A Novel Cytokine Derived Fusion Tag for Over- Expression of Heterologous Proteins in E. coli", 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", volume = "3", number = "3", pages = "157-5", }
