Abstract：
The occlusion bodies of Autographa californica multiple nucleopolyhedrovirus are proteinaceous formations with significant biotechnological potential owing to their capacity to integrate foreign proteins through fusion with polyhedrin, their primary component. However, the strategy for successful heterologous protein inclusion still requires further refinement. In this study, we conducted a comparative assessment of various conditions to achieve the embedding of recombinant proteins within polyhedra. Two baculoviruses were constructed: AcPHGFP (polh+), with GFP as a fusion to wild type (wt) polyhedrin and AcΔPHGFP (polh+), with GFP fused to a fragment corresponding to amino acids 19 to 110 of polyhedrin. These baculoviruses were evaluated by infecting Sf9 cells and stably transformed Sf9, Sf9POLH, and Sf9POLHE44G cells. The stably transformed cells contributed another copy of wt or a mutant polyhedrin, respectively. Polyhedra of each type were isolated and characterized by classical methods. The fusion PHGFP showed more-efficient incorporation into polyhedra than ΔPHGFP in the three cell lines assayed. However, ΔPHGFP polyhedron yields were higher than those of PHGFP in Sf9 and Sf9POLH cells. Based on an integral analysis of the studied parameters, it can be concluded that, except for the AcΔPHGFP/Sf9POLHE44G combination, deficiencies in one factor can be offset by improved performance by another. The combinations AcPHGFP/Sf9POLHE44G and AcΔPHGFP/Sf9POLH stand out due to their high level of incorporation and the large number of recombinant polyhedra produced, respectively. Consequently, the choice between these approaches becomes dependent on the intended application.
摘要：
加州自拟多核多角体病毒的闭塞体是蛋白质形成，具有显着的生物技术潜力，因为它们能够通过与多角体融合整合外源蛋白，他们的主要组成部分。然而,成功包含异源蛋白的策略仍需要进一步完善.在这项研究中,我们对各种条件进行了比较评估，以实现重组蛋白在多面体中的包埋。构建了两种杆状病毒：AcPHGFP（polh），将GFP与野生型（wt）多面体蛋白和ActΔPHGFP（polh）融合，与GFP融合到对应于多角体蛋白氨基酸19至110的片段。通过感染Sf9细胞和稳定转化的Sf9，Sf9POLH，和Sf9POLHE44G细胞。稳定转化的细胞贡献了另一个wt或突变多角体蛋白的拷贝，分别。分离出每种类型的多面体，并通过经典方法进行表征。在测定的三种细胞系中，融合PHGFP比ΔPHGFP更有效地掺入多面体。然而,在Sf9和Sf9POLH细胞中，ΔPHGFP多面体的产量高于PHGFP。在对所研究参数进行积分分析的基础上，可以得出结论，除ActΔPHGFP/Sf9POLHE44G组合外，一个因素的不足可以通过另一个因素的性能改进来弥补。AcPHGFP/Sf9POLHE44G和AcAPPHGFP/Sf9POLH的组合由于其高水平的掺入和产生的大量重组多面体而脱颖而出，分别。因此,这些方法之间的选择变得取决于预期的应用。
