Functional analysis of Spodoptera litura nucleopolyhedrovirus p49 gene during Autographa californica nucleopolyhedrovirus infection of SpLi-221 cells
Functional analysis of Spodoptera litura nucleopolyhedrovirus p49 gene during Autographa californica nucleopolyhedrovirus infection of SpLi-221 cells
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斜纹夜蛾核多角体病毒p49基因在苜蓿银纹夜蛾核多角体病毒感染SpLi-221细胞过程中的功能分析
DOI：
10.1007/s11262-010-0520-5
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发表时间：
2010-12-01
期刊：
影响因子：
1.6
通讯作者：
Pang, Yi
中科院分区：
文献类型：
--
作者：
Lin, Tiehao;Yu, Mei;Pang, Yi
斜纹夜蛾核多角体病毒 (SpltNPV) 抗凋亡基因 Splt-p49 能够挽救 p35 缺失的苜蓿银纹夜蛾核多角体病毒 (AcMNPV) 在 AcMNPV 许可的 Sf9 细胞中的复制。本研究通过大肠杆菌中的ET同源重组产生了AcMNPV p35敲除杆粒，命名为vAc(P35-KO)。将 Splt-p49 基因转置到 vAc(P35-KO) 的多角体基因座中，以研究 Splt-p49 在 AcMNPV 不容许的 SpLi-221 细胞感染 AcMNPV 的情况下是否具有任何抗凋亡活性。我们的结果表明，当 Splt-p49 在其天然启动子的控制下时，不能抑制 AcMNPV 感染 SpLi-221 细胞诱导的细胞凋亡。 Western blot分析显示Splt-P49表达较差。当Splt-P49的表达处于果蝇hsp70启动子的控制下时，Splt-P49的表达提前，并且检测到较高水平的Splt-P49。结果，SpLi-221细胞的凋亡受到抑制；然而，与 AcMNPV 感染的 SpLi-221 细胞相比，芽生病毒的产量并没有提高。这些数据表明，除了细胞凋亡之外，在不允许的 SpLi-221 细胞中，AcMNPV 复制还存在其他障碍。
The Spodoptera litura nucleopolyhedrovirus (SpltNPV) antiapoptotic gene Splt-p49 is able to rescue replication of a p35-null Autographa californica nucleopolyhedrovirus (AcMNPV) in AcMNPV-permissive Sf9 cells. In this study, an AcMNPV p35 knockout bacmid was generated through ET homologous recombination in Escherichia coli and designated as vAc(P35-KO). The Splt-p49 gene was transposed into the polyhedrin locus of vAc(P35-KO) to investigate if Splt-p49 has any antiapoptotic activity in the context of AcMNPV infection of AcMNPV-nonpermissive SpLi-221 cells. Our results demonstrated that Splt-p49 could not inhibit the apoptosis induced by AcMNPV infection of SpLi-221 cells when it was under the control of its native promoter. Western blot analysis showed that Splt-P49 was poorly expressed. When the expression of Splt-P49 was under the control of Drosophila hsp70 promoter, the expression of Splt-P49 was advanced, and a higher level of Splt-P49 was detected. As a result, the apoptosis of SpLi-221 cells was inhibited; however, budded-virus production did not improve in comparison with that in AcMNPV-infected SpLi-221 cells. These data indicated that there are other barrier(s) to AcMNPV replication in the nonpermissive SpLi-221 cells besides apoptosis.