Efficient expression of Human papillomavirus 16 E7 oncoprotein fused to C-terminus of Tobacco mosaic virus (TMV) coat protein using molecular chaperones in Escherichia coli.

Jitka Folwarczna, Tomas Moravec, Helena Plchova, Hana Hoffmeisterova, Noemi Cerovska

Institute of Experimental Botany, vvi, Academy of Sciences of the Czech Republic, Prague 6, Czech Republic.

Protein expression and purification 2012 Sep

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The Human papillomavirus 16 (HPV16) E7 oncoprotein is a promising candidate for development of anti-cancer therapeutic vaccine. We have prepared the expression construct carrying mutagenized E7 oncoprotein fused to the C-terminus of Tobacco mosaic virus (TMV) coat protein via 15 amino acids β-sheet linker. The fusion protein was expressed in Escherichia coli MC 1061 cells. We have obtained high level expression, but most of the protein remained in insoluble inclusion bodies. To increase the ratio of soluble protein various molecular chaperones (TF, DnaK-DnaJ-GrpE, GroEL-GroES) were used. The immunological reactivity of expressed recombinant protein was evaluated with anti-E7 and anti-TMV antibodies. The distribution of expressed product during ultracentrifugation on sucrose gradient was studied. Copyright © 2012 Elsevier Inc. All rights reserved.

Citation

Jitka Folwarczna, Tomas Moravec, Helena Plchova, Hana Hoffmeisterova, Noemi Cerovska. Efficient expression of Human papillomavirus 16 E7 oncoprotein fused to C-terminus of Tobacco mosaic virus (TMV) coat protein using molecular chaperones in Escherichia coli. Protein expression and purification. 2012 Sep;85(1):152-7