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Vol 47(2013) N 5 p. 733-742;
G.V. Kochneva1*, I.N. Babkina3, T.A. Lupan4, A.A. Grazhdantseva1, P.V. Yudin1, G.F. Sivolobova1, A.N. Shvalov4, E.G. Popov1, I.V. Babkin3, S.V. Netesov1, P.M. Chumakov2

Apoptin Enhances the Oncolytic Activity of Vaccinia Virus in vitro

1Novosibirsk State University, Novosibirsk, 630090 Russia
2Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, 119991 Russia
3Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of Russian Academy of Sciences, Novosibirsk, 630090 Russia
4State Research Center of Virology and Biotechnology "Vector", Koltsovo, 630559 Russia

*g.v.kochneva@yandex.ru
Received - 2013-03-31; Accepted - 2013-04-27

The chicken anemia virus gene that encodes apoptin, a selective killer of cancer cells, was synthesized and inserted into the vaccinia virus (strain L-IVP) genome. The insertion replaces a major part of the viral C11R gene that encodes viral growth factor, which is important for virulence. The recombinant virus VVdGF-ApoS24/2 was obtained by transient dominant selection using the gene of puromycin resistance as a selective marker. The expression of the apoptin gene from a synthetic early-late promoter of vaccinia virus ensured the efficient accumulation of the target protein in VVdGF-ApoS24/2-infected cells. Although recombinant apoptin carried the signal peptide of the virus growth factor at the N-end, the protein was not secreted into the culture medium. The recombinant virus VVdGF-ApoS24/2 exhibited significantly higher selective lytic activity in human cancer cell lines (A549, A431, U87MG, RD, and MCF7) than the parent strain L-IVP and its VVdGF2/6 variant with C11R deletion. These results suggest that the use of apoptin can be an efficient means of enhancing the natural anticancer activity of vaccinia virus.

apoptin, transient dominant selection, recombinant vaccinia virus, virus growth factor, puromycin, oncolytic activity



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