title: 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.
abstract: 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.
authors: Institute of Experimental Botany, vvi, Academy of Sciences of the Czech Republic, Prague 6, Czech Republic.
authors: Jitka
authors: J
authors: Folwarczna
authors: Tomas
authors: T
authors: Moravec
authors: Helena
authors: H
authors: Plchova
authors: Hana
authors: H
authors: Hoffmeisterova
authors: Noemi
authors: N
authors: Cerovska
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology; D036022:Capsid Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D003001:Cloning, Molecular / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D052162:Human papillomavirus 16 / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D006801:Humans; D051379:Mice; D018832:Molecular Chaperones / Q000737:chemistry / Q000235:genetics; D016296:Mutagenesis; D050725:Papillomavirus E7 Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D030361:Papillomavirus Infections / Q000821:virology; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D012995:Solubility; D014027:Tobacco Mosaic Virus / Q000737:chemistry / Q000235:genetics* / Q000276:immunology
chemical_list: D000906:Antibodies; D036022:Capsid Proteins; D018832:Molecular Chaperones; D050725:Papillomavirus E7 Proteins; D011993:Recombinant Fusion Proteins