Folwarczna J., Moravec T., Plchova H., Hoffmeisterova H., Cerovska N.
PROTEIN EXPRESSION AND PURIFICATION
85:
152-157,
2012
Klíčová slova:
bacterial expression, Human papillomavirus, E7 oncoprotein, chaperone
Abstrakt:
The Human papillomavirus 16 (HPV 16) 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.
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Autoři z ÚEB: Noemi Čeřovská,
Jitka Folwarczna,
Hana Hoffmeisterová,
Tomas Moravec,
Helena Plchová
Čeština
English