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

0380787 - ÚEB 2013 RIV US eng J - Článek v odborném periodiku
Folwarczna, Jitka - Moravec, Tomáš - Plchová, Helena - Hoffmeisterová, Hana - Čeřovská, Noemi
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. Roč. 85, č. 1 (2012), s. 152-157. ISSN 1046-5928. E-ISSN 1096-0279
Grant CEP: GA ČR GA521/09/1525; GA ČR GAP501/12/1761
Výzkumný záměr: CEZ:AV0Z50380511
Klíčová slova: Bacterial expression * Human papillomavirus * E7 oncoprotein
Kód oboru RIV: EI - Biotechnologie a bionika
Impakt faktor: 1.429, rok: 2012

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.
Trvalý link: http://hdl.handle.net/11104/0211407