Polyhedron envelope protein (PEP) is the major component of the calyx that surrounds the baculovirus occlusion body (OB). PEP has been associated with the stabilization and resistance of polyhedra in the environment. Due to the abundant levels of PEP in OBs, we decided to use this protein as a fusion partner to redirect foreign proteins to baculovirus polyhedra. In this study we developed a strategy that involves the generation of a monoclonal transformed insect cell line expressing a protein of interest fused to the the Anticarsia gemmatalis multiple nucleopolyhedrovirus (AgMNPV) N-terminus of PEP that enables the packaging of foreign proteins into the OBs without generating a recombinant baculovirus. This proved to be an efficient platform that could be exploited to improve wild type baculovirus for their use as bioinsecticides without facing the concerns of releasing genetically modified DNA to the environment and bypassing the associated regulatory issues. We demonstrated, using immunological, proteomic and microscopy techniques, that the envelope of AgMNPV OBs can effectively trap chimeric proteins in an infected insect cell line expressing AgMNPV PEP fused to the enhanced green fluorescent protein (eGFP). Furthermore, packaging of chimeric PEP also took place with heterologous OBs such as those of Autographa californica multiple nucleopolyhedrovirus (AcMNPV), another group I alphabaculovirus.

中文翻译：

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将外源蛋白质包装到杆状病毒闭塞体中而不对病毒基因组进行改造的简化策略。

多面体包膜蛋白（PEP）是花萼的主要成分，包围杆状病毒闭塞体（OB）。PEP与环境中多面体的稳定性和抗性有关。由于OB中的PEP含量很高，我们决定使用此蛋白作为融合伴侣，将外源蛋白重定向至杆状病毒多面体。在这项研究中，我们开发了一种策略，该策略涉及生成表达与PEP的抗癌双子核多角体病毒（AgMNPV）N末端融合的目标蛋白的单克隆转化昆虫细胞系，该方法能够将外源蛋白包装到OB中而无需产生重组杆状病毒。事实证明，这是一个有效的平台，可用于改进野生型杆状病毒作为生物杀虫剂的用途，而无需担心将转基因的DNA释放到环境中并绕开相关的监管问题。我们使用免疫学，蛋白质组学和显微镜技术证明，AgMNPV OB的包膜可以有效地将嵌合蛋白捕获在表达AgMNPV PEP并融合了增强绿色荧光蛋白（eGFP）的被感染昆虫细胞系中。此外，嵌合PEP的包装也与异源OB一起进行，所述异源OB如加利福尼亚州的Autographa californica多核多角体病毒（AcMNPV），另一类I杆状杆状病毒。使用免疫学，蛋白质组学和显微镜技术，AgMNPV OB的包膜可以有效地将嵌合蛋白捕获在表达AgMNPV PEP并与增强型绿色荧光蛋白（eGFP）融合的感染昆虫细胞系中。此外，嵌合PEP的包装也与异源OB一起进行，所述异源OB如加利福尼亚州的Autographa californica多核多角体病毒（AcMNPV），另一类I杆状杆状病毒。使用免疫学，蛋白质组学和显微镜技术，AgMNPV OB的包膜可以有效地将嵌合蛋白捕获在表达AgMNPV PEP并与增强型绿色荧光蛋白（eGFP）融合的感染昆虫细胞系中。此外，嵌合PEP的包装也与异源OB一起进行，所述异源OB如加利福尼亚州的Autographa californica多核多角体病毒（AcMNPV），另一类I杆状杆状病毒。