Effects of Early or Overexpression of the Autographa californica Multiple Nucleopolyhedrovirus orf94 (ODV-e25) on Virus Replication.
Effects of Early or Overexpression of the Autographa californica Multiple Nucleopolyhedrovirus orf94 (ODV-e25) on Virus Replication.
复制标题
苜蓿银纹夜蛾多重核多角体病毒 orf94 (ODV-e25) 早期或过度表达对病毒复制的影响
DOI：
10.1371/journal.pone.0065635
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Li LL
中科院分区：
文献类型：
--
作者：
Luo XC;Wang SS;Zhang J;Qian DD;Wang SM;Li LL
odv-e25（e25）是杆状病毒的核心基因之一。为了研究它如何在杆状病毒的复制周期中发挥作用，使用杆状病毒系统构建了许多具有e25的苜蓿银纹夜蛾多核型多角体病毒重组体，其在立即早期基因ie 1的启动子或极晚期超表达基因p10的启动子的控制下，并评估了e25的早期表达或过表达对病毒复制的影响。显微镜和滴定分析表明，在ie 1启动子控制下的e25的杆状病毒不能产生芽殖病毒，而在p10启动子控制下的e25的重组病毒产生芽殖病毒正常，但在感染细胞中的闭塞体的形成显着减少和延迟。电子显微镜观察显示，在ie 1启动子控制下的e25重组杆粒转染的细胞中没有成熟的病毒粒子或完整的核衣壳。实时定量PCR分析表明，e25启动子的改变不影响病毒DNA的合成。通过e25的早期表达，来自主要衣壳蛋白基因vp 39的启动子的报告基因表达降低了63%。共聚焦显微镜显示，E25主要定位在细胞核中的24小时后感染野生型病毒，但它仍然在细胞质中的重组杆粒转染与e25下的ie 1启动子的控制下，表明E25的运输到细胞核中的特定和严格的时间依赖性的方式进行调节。
odv-e25(e25) is one of the core genes of baculoviruses. To investigate how it functions in the replication cycle of a baculovirus, a number of Autographa californica multiple nucleopolyhedrovirus recombinants with e25 under control of the promoter of immediate early gene ie1, or the promoter of the very late hyperexpressed gene p10, were constructed using a bacmid system, and the effects of early expression or overexpression of e25 on replication of the virus were evaluated. Microscopy and titration assays demonstrated that bacmids with e25 under control of ie1 promoter were unable to produce budded viruses; and that the recombinant viruses with e25 under control of p10 promoter generated budded virus normally, but formation of occlusion bodies were dramatically reduced and delayed in the infected cells. Electron microscopy showed that there were no mature virions or intact nucleocapsids present in the cells transfected with a recombinant bacmid with e25 under control of ie1 promoter. Quantitative real-time PCR analysis demonstrated that alteration of the e25 promoter did not affect viral DNA synthesis. The reporter gene expression from the promoter of the major capsid protein gene vp39 was reduced 63% by early expression of e25. Confocal microscopy revealed that E25 was predominantly localized in nuclei by 24 hours post infection with wild-type virus, but it remained in the cytoplasm in the cells transfected with a recombinant bacmid with e25 under control of the ie1 promoter, suggesting that the transport of E25 into nuclei was regulated in a specific and strict time dependent manner.
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DOI：
10.1073/pnas.0402727101
发表时间：
2004-06-01
影响因子：
11.1
作者：
Braunagel, SC;Williamson, ST;Summers, MD
通讯作者：
Summers, MD
DOI：
10.1073/pnas.120163297
发表时间：
2000-06-06
影响因子：
11.1
作者：
Datsenko, KA;Wanner, BL
通讯作者：
Wanner, BL
影响因子：
3.7
作者：
KOVACS, GR;GUARINO, LA;SUMMERS, MD
通讯作者：
SUMMERS, MD
影响因子：
3.7
作者：
Beniya, H;Braunagel, SC;Summers, MD
通讯作者：
Summers, MD
影响因子：
5.4
作者：
CHAABIHI, H;OGLIASTRO, MH;CERUTTI, M
通讯作者：
CERUTTI, M