Investigation of the impact of cowpox virus BTB/kelch gene deletion on some characteristics of infection in vitro
2009
Kochneva, G.V. | Taranov, O.S. | Lupan, T.A. | Yudin, P.V. | Ryabchikova, E.I. | Drosdov, I.G. | Kolosova, I.A. | Shchelkunov, S.N., State Research Center of Virology and Biotechnology, Novosibirsk Region (Russian Federation) | Pubtsov, N.B. | Bajborodin, S.I., Russian Academy of Sciences. Siberian Branch, Novosibirsk (Russian Federation). Institute of Cytology and Genetics
A feature of reproduction of cowpox virus (CPV) mutants with target deletion of 4 of the BTB (broad complex, tramtrack and brick a brack)/kelch genes (D11L, C18L, G3L and A57R) was examined in CV-1 cell cultures, reorganization of actinic cytoskeleton in particular. The work involved wild-type CPV (WT CPV) and mutants constructed on its basis 216 and 337 with deletion of D11, G3L, C18L and A57R BTB/kelch genes. The study has shown a relationship of BTB/kelch genes and temperature sensitivity of CPV. There were no changes WT CPV and mutants 216 and 337. But during 72-h incubation the production of quarterly mutants was as much as 100times lower compared to WT CPV. The reduced cytopathic effect promoted longer preservation of infected cells and increased the reproduction period of virus. A less number of cell with virus-induced cytoplasmatic protrusions (63 cells of infected 300) was formed in the cell culture CV-1 infected with mutants compared to WT CPV (127 of 300). Confocal microscopy revealed the formation of large globed structures in the cells infected with mutants that were recognized as incomplete protrusions comprising virus antigens and actin. Taking into consideration that the protrusions contained mature virus particles, it is concluded that the formation of long protrusions with infected cells is one of mechanisms of spreading poxvirus infection in vitro. The activity of virus kelch-genes partially compensates destroying the actinic cytoskeleton of an infected cell in the reproduction of poxviruses providing movement of virus particles at great distances from the cell body.
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