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在允许性、半允许性和非允许性细胞系感染期间,对选定的甜菜夜蛾多核型多角体病毒基因启动子进行实时表达分析。

Real-Time Expression Analysis of Selected Anticarsia gemmatalis multiple nucleopolyhedrovirus Gene Promoters during Infection of Permissive, Semipermissive and Nonpermissive Cell Lines.

作者信息

Morgado Fabricio da Silva, Ardisson-Araújo Daniel Mendes Pereira, Ribeiro Bergmann Morais

机构信息

Laboratory of Baculovirus, Cell Biology Department, University of Brasília, 70910-900 Brasília-DF, Brazil.

Laboratory of Insect Virology, Department of Biochemistry and Molecular Biology, Federal University of Santa Maria, 97105-900 Santa Maria-RS, Brazil.

出版信息

Viruses. 2017 Jun 1;9(6):132. doi: 10.3390/v9060132.

Abstract

Baculovirus infection follows a transcriptionally controlled sequence of gene expression that occurs by activation of different viral gene promoter sequences during infection. This sequence of promoter activation may be disrupted by cellular defenses against viral infection, which might interfere with viral progeny formation. In this work, the activity of the , , , , and promoters of the was assessed during infection of permissive, semipermissive and nonpermissive cell lines by a novel methodology that detects reporter protein luminescence in real-time. This technique allowed us to characterize in rich detail the AgMNPV promoters in permissive cell lines and revealed differential profiles of expression in cells with limited permissivity that correlate well with limitations in viral DNA replication. Semipermissive and nonpermissive cell lines presented delays and restrictions in late and very late promoter expression. Cells undergoing apoptosis did not inhibit late gene expression; however, viral progeny formation is severely affected. This work demonstrates the application of the real-time luminescence detection methodology and how the promoter expression profile may be used to diagnose cellular permissivity to baculovirus infection.

摘要

杆状病毒感染遵循基因表达的转录控制序列,该序列在感染期间通过激活不同的病毒基因启动子序列而发生。启动子激活的这一序列可能会被细胞针对病毒感染的防御机制破坏,这可能会干扰病毒子代的形成。在这项工作中,通过一种实时检测报告蛋白发光的新方法,评估了苜蓿银纹夜蛾核型多角体病毒(AgMNPV)的、、、、和启动子在允许性、半允许性和非允许性细胞系感染过程中的活性。这项技术使我们能够详细地描述允许性细胞系中AgMNPV启动子的特征,并揭示了在允许性有限的细胞中不同的表达谱,这与病毒DNA复制的限制密切相关。半允许性和非允许性细胞系在晚期和极晚期启动子表达上出现延迟和限制。正在经历凋亡的细胞不会抑制晚期基因表达;然而,病毒子代的形成受到严重影响。这项工作展示了实时发光检测方法的应用,以及启动子表达谱如何可用于诊断细胞对杆状病毒感染的允许性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d31a/5490809/b70a49c9e60c/viruses-09-00132-g001.jpg

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