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首页> 外文期刊>Applied Microbiology and Biotechnology >Expression of the lef5 gene from Spodoptera exigua multiple nucleopolyhedrovirus contributes to the baculovirus stability in cell culture
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Expression of the lef5 gene from Spodoptera exigua multiple nucleopolyhedrovirus contributes to the baculovirus stability in cell culture

机译：来自Spodoptera exigua的Lef5基因的表达多核多核病毒有助于细胞培养的杆状病毒稳定性

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Baculoviruses are a broad group of viruses infecting insects, predominately of the order Lepidoptera. They are used worldwide as biological insecticides and as expression vectors to produce recombinant proteins. Baculoviruses replicate in their host, although several cell lines have been developed for in vitro replication. Nevertheless, replication of baculoviruses in cell culture involves the generation of defective viruses with a decrease in productivity and virulence. Transcriptional studies of the Spodoptera exigua multiple nucleopolyhedrovirus (SeMNPV) and the Autographa californica multiple nucleopolyhedrovirus (AcMNPV) infective process revealed differences in the expression patterns when the virus replicated under in vitro (Se301 cells) or in vivo (S. exigua larvae) conditions. The late expression factor 5 (lef5) gene was found to be highly overexpressed when the virus replicates in larvae. To test the possible role of lef5 expression in viral stability, recombinant AcMNPV expressing the lef5 gene from SeMNPV (Se-lef5) was generated and its stability was monitored during successive infection passages in Sf21 cells by evaluating the loss of several essential and non-essential genes. The gfp transgene was more stable in those viruses expressing the Se-LEF5 protein and the GFP-defective viruses were accumulated at a lower level when compared to its control viruses, confirming the positive influence of lef5 in viral stability during the multiplication process. This work describes for the first time a viral factor involved in transgene stability when baculoviruses replicate in cell culture, opening new ways to facilitate the in vitro production of recombinant proteins using baculovirus.
机译：Baculoviruses是一种感染昆虫的广泛病毒，主要是鳞翅目的命令。它们作为生物杀虫剂和作为制备重组蛋白的表达载体使用。杆状病毒在其宿主中复制，尽管已经开发了用于体外复制的几种细胞系。然而，细胞培养中的杆状病毒的复制涉及产生有缺陷的病毒，随着生产率和毒力而降低。 Spodoptera exigua多核多发性rovirus（semnpv）的转录研究和术语加州多核心子病毒（Acmnpv）感染过程揭示了当在体外（SE301细胞）或体内（S. exigua幼虫）条件下的病毒时表达模式的差异。当病毒在幼虫复制时，发现晚表达因子5（LEF5）基因被发现高度过度表达。为了测试LeF5表达在病毒稳定性中的可能作用，产生从SEMNPV（SE-LEF5）的重组ACMNPV，通过评估几种必要和非必要的丧失，在SF21细胞中的连续感染通道期间监测其稳定性基因。在表达SE-LEF5蛋白的那些病毒中，GFP转基因更稳定，与其对照病毒相比，GFP缺陷病毒在较低水平上累积，确认在乘法过程中LEF5在病毒稳定性中的阳性影响。这项工作描述了第一次参与转基因稳定性的病毒因素当杆状病毒在细胞培养中复制时，开启新方法以促进使用杆状病毒的重组蛋白的体外产生。

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