1887

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Volume 89, Issue 8

Functional analysis of the putative antiapoptotic genes, and , of nucleopolyhedrovirus with RNAi Free

Abstract

A homology search of a public database revealed that nucleopolyhedrovirus (SpltNPV) possesses two putative, antiapoptotic genes, and inhibitor of apoptosis 4 (), but their function has not been investigated in its native host cells. In the present study, we used RNA interference (RNAi) to silence the expression of and , independently or together, to determine their roles during the SpltNPV life cycle. RT-PCR analysis and Western blot analysis showed the target gene expression had been knocked out in the SpltNPV-infected SpLi-221 cells after treatment with or double-stranded RNA (dsRNA), respectively, confirming that the two genes were effectively silenced. In SpltNPV-infected cells treated with dsRNA, apoptosis was observed beginning at 14 h, and almost all cells had undergone apoptosis by 48 h. In contrast, budded virus production and polyhedra formation progressed normally in infected cells treated with dsRNA. Cell viability analysis showed that Splt-IAP4 had no synergistic effect on the inhibition of apoptosis of SpLi-221 cells induced by SpltNPV infection. Interestingly, after dsRNA treatment, cells did not congregate like those infected with SpltNPV in the early infection phase, implying an unknown role of baculovirus . Our results determine that is necessary to prevent apoptosis; however, has no antiapoptotic function during SpltNPV infection.

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2008-08-01
2024-04-19
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Functional analysis of the putative antiapoptotic genes, p49 and iap4, of Spodoptera litura nucleopolyhedrovirus with RNAi
J. Gen. Virol. 89, 1873 (2008); https://doi.org/10.1099/vir.0.2008/001008-0
/content/journal/jgv/10.1099/vir.0.2008/001008-0
/content/journal/jgv/10.1099/vir.0.2008/001008-0
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