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

Folwarczna, Jitka; Moravec, Tomáš; Plchová, Helena; Hoffmeisterová, Hana; Čeřovská, Noemi

doi:https://dx.doi.org/10.1016/j.pep.2012.07.008

Number of the records: 1

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

1.

SYSNO ASEP	0380787
Document Type	J - Journal Article
R&D Document Type	Journal Article
Subsidiary J	Článek ve WOS
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
Author(s)	Folwarczna, Jitka (UEB-Q) Moravec, Tomáš (UEB-Q)_{RID, ORCID} Plchová, Helena (UEB-Q)_{RID, ORCID} Hoffmeisterová, Hana (UEB-Q)_RID Čeřovská, Noemi (UEB-Q)_{RID, ORCID}
Source Title	Protein Expression and Purification. - : Elsevier - ISSN 1046-5928 Roč. 85, č. 1 (2012), s. 152-157
Number of pages	6 s.
Language	eng - English
Country	US - United States
Keywords	Bacterial expression ; Human papillomavirus ; E7 oncoprotein
Subject RIV	EI - Biotechnology ; Bionics
R&D Projects	GA521/09/1525 GA ČR - Czech Science Foundation (CSF)
	GAP501/12/1761 GA ČR - Czech Science Foundation (CSF)
CEZ	AV0Z50380511 - UEB-Q (2005-2011)
UT WOS	000308630400021
DOI	10.1016/j.pep.2012.07.008
Annotation	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.
Workplace	Institute of Experimental Botany
Contact	David Klier, knihovna@ueb.cas.cz, Tel.: 220 390 469
Year of Publishing	2013

Number of the records: 1