Protein Expression Laboratory, NCI RAS Initiative, Cancer Research Technology Program, Frederick National Laboratory for Cancer Research, Frederick, MD, 21702, USA.
Protein Expression Laboratory, NCI RAS Initiative, Cancer Research Technology Program, Frederick National Laboratory for Cancer Research, Frederick, MD, 21702, USA.
Protein Expr Purif. 2021 Oct;186:105924. doi: 10.1016/j.pep.2021.105924. Epub 2021 Jun 1.
Generating recombinant proteins in insect cells has been made possible via the use of the Baculovirus Expression Vector System (BEVS). Despite the success of many proteins via this platform, some targets remain a challenge due to issues such as cytopathic effects, the unpredictable nature of co-infection and co-expressions, and baculovirus genome instability. Many promoters have been assayed for the purpose of expressing diverse proteins in insect cells, and yet there remains a lack of implementation of those results when reviewing the landscape of commercially available baculovirus vectors. In advancing the platform to produce a greater variety of proteins and complexes, the development of such constructs cannot be avoided. A better understanding of viral gene regulation and promoter options including viral, synthetic, and insect-derived promoters will be beneficial to researchers looking to utilize BEVS by recruiting these intricate mechanisms of gene regulation for heterologous gene expression. Here we summarize some of the developments that could be utilized to improve the expression of recombinant proteins and multi-protein complexes in insect cells.
利用杆状病毒表达载体系统(BEVS)可以在昆虫细胞中生成重组蛋白。尽管通过该平台已经成功表达了许多蛋白质,但由于细胞病变效应、共感染和共表达的不可预测性以及杆状病毒基因组不稳定性等问题,某些靶标仍然具有挑战性。已经对许多启动子进行了检测,目的是在昆虫细胞中表达不同的蛋白质,但在回顾商业可用杆状病毒载体的情况时,这些结果的实施仍然缺乏。为了在该平台上生产更多种类的蛋白质和复合物,这些构建体的开发是不可避免的。更好地了解病毒基因调控和启动子选择,包括病毒、合成和昆虫衍生的启动子,将有助于研究人员通过招募这些复杂的基因调控机制来利用 BEVS 进行异源基因表达。在这里,我们总结了一些可以用来提高昆虫细胞中重组蛋白和多蛋白复合物表达的进展。