文献类型：期刊文献

英文题名：YBac: A novel and smple baculovirus system for heterologous protein production in Spodoptera frugiperda 9 (Sf9) cells

作者：Yu, Yue[1];Yang, Cancan[1];Zhang, Tong[1];Xu, Zhenhe[1];Zhang, Yuanxing[2];Liu, Qin[1,2,3]

机构：[1]East China Univ Sci & Technol, Shanghai Frontiers Sci Ctr Optogenet Tech Cell Met, State Key Lab Bioreactor Engn, Shanghai, Peoples R China;[2]Shanghai Collaborat Innovat Ctr Biomfg, Shanghai, Peoples R China;[3]East China Univ Sci & Technol, Shanghai Frontiers Sci Ctr Optogenet Tech Cell Met, State Key Lab Bioreactor Engn 1, Shanghai, Peoples R China

年份：2023

卷号：18

期号：7

外文期刊名：BIOTECHNOLOGY JOURNAL

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000979799700001)】；

基金：This work was supported by Shanghai Frontiers Science Center of Optogenetic Techniques for Cell Metabolism grant 2021 Sci & Tech 03-28 (Shanghai Municipal Education Commission) and The Fundamental Research Funds for the Central Universities (222201211716).

语种：英文

外文关键词：baculovirus; gene deletion; heterologous protein expression; homologous recombination; insect cells

摘要：The baculovirus expression vector system (BEVS) has been widely used for heterologous protein expression due to its powerful functionality and easy access to commercial expression vectors. Currently, most laboratories prefer two strategies for protein production using BEVS. One is recombinant bacmid based on transposition in Escherichia coli (e.g., Bac-to-Bac), and another is homologous recombination in insect cells (e.g., flashBac). In this manuscript, a rapid and simple YBac system was established. This novel system uses an Ac99KO bacmid as a virus vector and co-transfected into Spodoptera frugiperda 9 (Sf9) cells with a donor plasmid capable of recombination into Ac42 loci that carry the genes of interest (GOIs) along with the complete Ac99 fragment. Based on the intracellular homologous recombination system, the production of foreign proteins was achieved by the complementation of the Ac99 gene together with the insertion of GOIs. In this study, the human thyroid peroxidase (hTPO) and porcine epidemic diarrhea virus-like particles (PEDV VLPs) were successfully expressed using the YBac system. The entire process was shortened to 10 days, and the components involved in the system could be easily prepared in the laboratory, suggesting that the YBac system may have great potential in the production of heterologous proteins.